KRAS G12C inhibitors and methods of using the same

ABSTRACT

Provided herein are KRAS G12C inhibitors, such as 
     
       
         
         
             
             
         
       
         
         
           
             composition of the same, and methods of using the same. These inhibitors are useful for treating a number of disorders, including pancreatic, colorectal, and lung cancers.

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional patentapplication 62/679,655 filed on Jun. 1, 2018, which specification ishereby incorporated herein by reference in its entirety for allpurposes.

FIELD OF THE INVENTION

The present invention relates to compounds that inhibit the KRAS G12Cprotein; methods of treating diseases or conditions, such as cancer,using the compounds; and pharmaceutical compositions containing thecompounds.

BACKGROUND

KRAS gene mutations are common in pancreatic cancer, lungadenocarcinoma, colorectal cancer, gall bladder cancer, thyroid cancer,and bile duct cancer. KRAS mutations are also observed in about 25% ofpatients with NSCLC, and some studies have indicated that KRAS mutationsare a negative prognostic factor in patients with NSCLC. Recently,V-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutationshave been found to confer resistance to epidermal growth factor receptor(EGFR) targeted therapies in colorectal cancer; accordingly, themutational status of KRAS can provide important information prior to theprescription of TKI therapy. Taken together, there is a need for newmedical treatments for patients with pancreatic cancer, lungadenocarcinoma, or colorectal cancer, especially those who have beendiagnosed to have such cancers characterized by a KRAS mutation, andincluding those who have progressed after chemotherapy.

The compounds disclosed herein can be in the form of a pharmaceuticallyacceptable salt. The compounds provided can be formulated into apharmaceutical formulation comprising a compound disclosed herein and apharmaceutically acceptable excipient.

Also provided is a method of inhibiting KRAS G12C in a cell, comprisingcontacting the cell with a compound or composition disclosed herein.Further provided is a method of treating cancer in a subject comprisingadministering to the subject a therapeutically effective amount of acompound or composition disclosed herein. In some embodiments, thecancer is lung cancer, pancreatic cancer, or colorectal cancer.

SUMMARY

In one aspect of the present invention, the invention provides acompound having a structure of formula (I):

-   -   wherein        -   R¹ is a —C₁-C₆ alkyl, or —C₃-C₆cycloalkyl, group;        -   R^(1a) is a —C₁-C₆ alkyl, —C₁-C₆ heteroalkyl, aryl,            heteroaryl, —C₃-C₆ cycloalkyl or —C₃-C₆ heterocycloalkyl            group;        -   R¹ and R^(1a) together with the carbon atom to which they            are attached, form a carbocyclic ring, wherein the            carbocyclic ring can be unsubstituted or fused to an            aromatic ring;        -   R² is an aryl substituted with a halo, —OH, or NH₂;        -   R³ is halo;        -   R⁴ is H or methyl;        -   R⁴ is H or methyl; or    -   a stereoisomer thereof, an atropisomer thereof, a        pharmaceutically acceptable salt thereof, a pharmaceutically        acceptable salt of the stereoisomer thereof, or a        pharmaceutically acceptable salt of the atropisomer thereof.

One aspect of the present invention provides various compounds,stereoisomers, atropisomers, pharmaceutically acceptable salts,pharmaceutically acceptable salts of the stereoisomers, andpharmaceutically acceptable salts of the atropisomers as described inthe embodiments set forth below.

Another aspect of the present invention provides a pharmaceuticalcomposition that includes the compound of any of the embodiments or thepharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier or diluent.

Another aspect of the present invention provides a method of treatingcancer. Such methods include: administering to a patient in need thereofa therapeutically effective amount of the compound of any of theembodiments or a pharmaceutically acceptable salt thereof. In some suchmethods, the cancer is a hematologic malignancy. In some such methods,the cancer is selected from the group consisting of breast cancer,colorectal cancer, skin cancer, melanoma, ovarian cancer, kidney cancer,lung cancer, non-small cell lung cancer, lymphoma, non-Hodgkin'slymphoma, myeloma, multiple myeloma, leukemia, and acute myelogenousleukemia. In some other such methods, the cancer is multiple myeloma. Insome other such methods, the cancer is acute myelogenous leukemia. Insome other such methods, the cancer is non-Hodgkin's lymphoma.

In another aspect, the method further includes administering to apatient in need thereof a therapeutically effective amount of one ormore additional pharmaceutically active compounds. For example, in somesuch methods the one or more additional pharmaceutically activecompounds is carfilzomib. In others, the one or more additionalpharmaceutically active compounds is venetoclax. In still other suchmethods, the one or more additional pharmaceutically active compounds iscytarabine. In still other such methods, the one or more additionalpharmaceutically active compounds is daratumumab. In still other suchmethods, the one or more additional pharmaceutically active compounds isan MCI-1 inhibitor. In still other such methods, the MCI-1 inhibitor isAMG-176. In still other such methods, the one or more additionalpharmaceutically active compounds is an immunomodulatory iMID.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure belongs. Methods and materials aredescribed herein for use in the present disclosure; other, suitablemethods and materials known in the art can also be used. The materials,methods, and examples are illustrative only and not intended to belimiting. All publications, patent applications, patents, sequences,database entries, and other references mentioned herein are incorporatedby reference in their entirety. In case of conflict, the presentspecification, including definitions, will control.

Other features and advantages of the disclosure will be apparent fromthe following detailed description and figures, and from the Claims.

DETAILED DESCRIPTION Definitions

Abbreviations: The following abbreviations may be used herein:

AcOH acetic acid aq or aq. aqueous BOC or Boc tert-butyloxycarbonyl DCMdichloromethane DIPEA or Hunig's Base N,N-diisopropylethylamine DMAP4-dimethylaminopyridine DME 1,2-dimethoxyethane DMFN,N-dimethylformamide DMSO dimethyl sulfoxide Dppf, DPPF or dppf1,1′-bis(diphenylphosphino)ferrocene eq or eq. or equiv. equivalent ESIor ES electrospray ionization Et ethyl Et₂O diethyl ether EtOAc ethylacetate g gram(s) h or hr hour(s) HPLC high pressure liquidchromatography iPr isopropyl iPr₂NEt or DIPEA N-ethyl diisopropylamine(Hünig's base) KHMDS potassium hexamethyldisilazide KOAc potassiumacetate LC MS, LCMS, liquid chromatography mass LC-MS or LC/MSspectroscopy LHMDS or LiHMDS lithium hexamethyldisilazide m/z massdivided by charge Me methyl MeCN acetonitrile MeOH methanol mgmilligrams min minutes mL milliliters MS mass spectra NaHMDS sodiumhexamethyldisilazide NBS N-bromosuccinimide n-BuLi n-butyllithium NCSN-chlorosuccinimide NMR nuclear magnetic resonance Pd₂(dba)₃tris(dibenzylideneacetone)dipalladium(0) Pd(dppf)Cl₂·DCM,[1,1′-bis(diphenylphosphino)ferrocene] Pd(dppf)Cl₂dichloropalladium(II), complex with dichloromethane Pd(PPH₃)₄tetrakis(triphenylphosphine)palladium(0) Ph phenyl PR or PG or Prot.group protecting group rbf round-bottom flask RP-HPLC reverse phase highpressure liquid chromatography RT or rt or r.t. room temperature sat, orsat'd saturated SFC supercritical fluid chromatography TBAFtetra-n-butylammonium fluoride TEA or Et₃N triethylamine TFAtrifluoroacetic acid THF tetrahydrofuran UV ultraviolet

The use of the terms “a,” “an,” “the,” and similar referents in thecontext of describing the invention (especially in the context of theclaims) are to be construed to cover both the singular and the plural,unless otherwise indicated. Recitation of ranges of values herein merelyare intended to serve as a shorthand method of referring individually toeach separate value falling within the range, unless otherwise indicatedherein, and each separate value is incorporated into the specificationas if it were individually recited herein. The use of any and allexamples, or exemplary language (e.g., “such as”) provided herein, isintended to better illustrate the invention and is not a limitation onthe scope of the invention unless otherwise claimed. No language in thespecification should be construed as indicating any non-claimed elementas essential to the practice of the invention.

As used herein, the term “alkyl” refers to straight chained and branchedC1-C₈ hydrocarbon groups, including but not limited to, methyl, ethyl,npropyl, ipropyl, nbutyl, secbutyl, tbutyl npentyl, 2methylbutyl,3methylbutyl, 2,2dimethylpropyl, nhexyl, 2methylpentyl, 3-methylpentyl,4-methylpentyl, 2,2dimethylbutyl, 2,3dimethylbutyl, 3,3dimethylbutyl,and 2ethybutyl. The term C_(m-n) means the alkyl group has “m” to “n”carbon atoms. The term “alkylene” refers to an alkyl group having asubstituent. An alkyl (e.g., methyl), or alkylene (e.g., —CH₂—), groupcan be substituted with one or more, and typically one to three, ofindependently selected, for example, halo, trifluoromethyl,trifluoromethoxy, hydroxy, alkoxy, nitro, cyano, alkylamino, C₁₋₆alkyl,C₂₋₆alkenyl, C₂₋₆alkynyl, —NC, amino, —CO₂H, —CO₂C₁-C₆alkyl,—OCOC₁-C₆alkyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ heterocycloalkyl, C₅-C₁₀aryl,and C₅-C₁₀ heteroaryl. The term “haloalkyl” specifically refers to analkyl group wherein at least one, e.g., one to six, or all of thehydrogens of the alkyl group are substituted with halo atoms.

The terms “alkenyl” and “alkynyl” indicate an alkyl group that furtherincludes a double bond or a triple bond, respectively.

As used herein, the term “halo” refers to fluoro, chloro, bromo, andiodo. The term “alkoxy” is defined as —OR, wherein R is alkyl.

As used herein, the term “amino” or “amine” interchangeably refers to a—NR₂ group, wherein each R is, e.g., H or a substituent. In someembodiments, the amino group is further substituted to form an ammoniumion, e.g., NR₃ ⁺. Ammonium moieties are specifically included in thedefinition of “amino” or “amine.” Substituents can be, for example, analkyl, alkoxy, cycloalkyl, heterocycloalkyl, amide, or carboxylate. An Rgroup may be further substituted, for example, with one or more, e.g.,one to four, groups selected from halo, cyano, alkenyl, alkynyl, alkyl,cycloalkyl, heterocycloalkyl, aryl, heteroaryl, urea, carbonyl,carboxylate, amine, and amide. An “amide” or “amido” groupinterchangeably refers to a group similar to an amine or amino group butfurther including a C(O), e.g., —C(O)NR₂. Some contemplated amino oramido groups (some with optional alkylene groups, e.g., alkylene-amino,or alkylene-amido) include CH₂NH₂, CH(CH₃)NH₂, CH(CH₃)₂NH₂, CH₂CH₂NH₂,CH₂CH₂N(CH₃)₂, CH₂NHCH₃, C(O)NHCH₃, C(O)N(CH₃)₂, CH₂C(O)NHphenyl,CH₂NHC(O)CH₃, CH₂NHCH₂OH, CH₂NHCH₂CO₂H, CH₂NH(CH₃)CH₂CO₂CH₃,CH₂NHCH₂CH₂OCH₃, CH₂NH(CH₃)CH₂CH₂OCH₃, CH₂NH(CH₃)CH₂C(O)N(CH₃)₂,CH₂NH(CH₃)CH₂C(O)NHCH₃, CH₂CH₂CCH, CH₂NMe₂, CH₂NH(CH₃)CH₂CH₂OH,CH₂NH(CH₃)CH₂CH₂F, CH₂N⁺(CH₃)₃, CH₂NHCH₂CHF₂, CH₂NHCH₂CH₃,

As used herein, the term “aryl” refers to a C₆₋₁₄ monocyclic orpolycyclic aromatic group, preferably a C₆₋₁₀ monocyclic or bicyclicaromatic group, or C₁₀₋₁₄, polycyclic aromatic group. Examples of arylgroups include, but are not limited to, phenyl, naphthyl, fluorenyl,azulenyl, anthryl, phenanthryl, pyrenyl, biphenyl, and terphenyl. Arylalso refers to C₁₀₋₁₄ bicyclic and tricyclic carbon rings, where onering is aromatic and the others are saturated, partially unsaturated, oraromatic, for example, dihydronaphthyl, indenyl, indanyl, ortetrahydronaphthyl (tetralinyl). Unless otherwise indicated, an arylgroup can be unsubstituted or substituted with one or more, and inparticular one to four, groups independently selected from, for example,halo, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, —CF₃, —OCF₃, —NO₂, —CN, —NC,—OH, alkoxy, amino, —CO₂H, —CO₂C₁-C₆alkyl, —OCOC₁-C₆alkyl, C₃-C₁₀cycloalkyl, C₃-C₁₀ heterocycloalkyl, C₅-C₁₀aryl, and C₅-C₁₀ heteroaryl.

As used herein, the term “carbocyclic ring” refers to a monocyclic ringwhich only includes carbon atoms as ring members. Such rings may befully saturated, partially saturated, or aromatic and may include 3 to10 carbon atoms.

As used herein, the term “cycloalkyl” refers to a monocyclic orpolycyclic non-aromatic carbocyclic ring, where the polycyclic ring canbe fused, bridged, or spiro. The carbocyclic ring can have 3 to 10carbon ring atoms. Contemplated carbocyclic rings include, but are notlimited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl, and cyclononyl.

As used herein, the term “heterocycloalkyl” means a monocyclic orpolycyclic (e.g., bicyclic), saturated or partially unsaturated, ringsystem containing 3 or more (e.g., 3 to 12, 4 to 10, 4 to 8, or 5 to 7)total atoms, of which one to five (e.g., 1, 2, 3, 4, or 5) of the atomsare independently selected from nitrogen, oxygen, and sulfur.Nonlimiting examples of heterocycloalkyl groups include azetidinyl,pyrrolidinyl, piperidinyl, piperazinyl, dihydropyrrolyl, morpholinyl,thiomorpholinyl, dihydropyridinyl, oxacycloheptyl, dioxacycloheptyl,thiacycloheptyl, and diazacycloheptyl.

Unless otherwise indicated, a cycloalkyl or heterocycloalkyl group canbe unsubstituted or substituted with one or more, and in particular oneto four, groups. Some contemplated substituents include halo, C₁₋₆alkyl,C₂₋₆alkenyl, C₂₋₆alkynyl, —OCF₃, —NO₂, —CN, —NC, —OH, alkoxy, amino,—CO₂H, —CO₂C₁-C₆alkyl, —OCOC₁-C₈alkyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀heterocycloalkyl, C₅-C₁₀aryl, and C₅-C₁₀ heteroaryl.

As used herein, the term “heteroaryl” refers to a monocyclic orpolycyclic ring system (for example, bicyclic) containing one to threearomatic rings and containing one to four (e.g., 1, 2, 3, or 4)heteroatoms selected from nitrogen, oxygen, and sulfur in an aromaticring. In certain embodiments, the heteroaryl group has from 5 to 20,from 5 to 15, from 5 to 10 ring, or from 5 to 7 atoms. Heteroaryl alsorefers to C₁₀₋₁₄ bicyclic and tricyclic rings, where one ring isaromatic and the others are saturated, partially unsaturated, oraromatic. Examples of heteroaryl groups include, but are not limited to,furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl,pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl,thiadiazolyl, thiazolyl, thienyl, tetrazolyl, triazinyl, triazolyl,benzofuranyl, benzimidazolyl, benzoisoxazolyl, benzopyranyl,benzothiadiazolyl, benzothiazolyl, benzothienyl, benzothiophenyl,benzotriazolyl, benzoxazolyl, furopyridyl, imidazopyridinyl,imidazothiazolyl, indolizinyl, indolyl, indazolyl, isobenzofuranyl,isobenzothienyl, isoindolyl, isoquinolinyl, isothiazolyl,naphthyridinyl, oxazolopyridinyl, phthalazinyl, pteridinyl, purinyl,pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalinyl, quiazolinyl,thiadiazolopyrimidyl, and thienopyridyl. Unless otherwise indicated, aheteroaryl group can be unsubstituted or substituted with one or more,and in particular one to four or one or two, substituents. Contemplatedsubstituents include halo, C₁₋₆alkyl, C₂₋₆alkenyl, C₂₋₆alkynyl, —OCF₃,—NO₂, —CN, —NC, —OH, alkoxy, amino, —CO₂H, —CO₂C₁-C₆alkyl,—OCOC₁-C₆alkyl, C₃-C₁₀ cycloalkyl, C₃-C₁₀ heterocycloalkyl, C₅-C₁₀aryl,and C₅-C₁₀ heteroaryl.

As used herein, the term Boc refers to the structure

As used herein, the term Cbz refers to the structure

As used herein, the term Bn refers to the structure

As used herein, the term trifluoroacetamide refers to the structure

As used herein, the term trityl refers to the structure

As used herein, the term tosyl refers to the structure

As used herein, the term Troc refers to the structure

As used herein, the term Teoc refers to the structure

As used herein, the term Alloc refers to the structure

As used herein, the term Fmoc refers to the structure

Compounds of the Disclosure

The compounds disclosed herein include all pharmaceutically acceptableisotopically-labeled compounds wherein one or more atoms of thecompounds disclosed herein are replaced by atoms having the same atomicnumber, but an atomic mass or mass number different from the atomic massor mass number usually found in nature. Examples of isotopes that can beincorporated into the disclosed compounds include isotopes of hydrogen,carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine,such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹³N, ¹⁵N, ¹⁵O, ¹⁷O, ¹⁸O, ³¹P, ³²P, ³⁵S,¹⁸F, ³⁶C, ¹²³I, and ¹²⁵I, respectively. These radiolabelled compoundscould be useful to help determine or measure the effectiveness of thecompounds, by characterizing, for example, the site or mode of action,or binding affinity to pharmacologically important site of action.Certain isotopically-labeled compounds of the disclosure, for example,those incorporating a radioactive isotope, are useful in drug and/orsubstrate tissue distribution studies. The radioactive isotopes tritium,i.e. ³H, and carbon-14, i.e. ¹⁴C, are particularly useful for thispurpose in view of their ease of incorporation and ready means ofdetection.

Substitution with heavier isotopes such as deuterium, i.e. 2H, mayafford certain therapeutic advantages resulting from greater metabolicstability, for example, increased in vivo half-life or reduced dosagerequirements, and hence are preferred in some circumstances.

Substitution with positron emitting isotopes, such as ¹¹C, ¹⁸F, ¹⁵O and¹³N, can be useful in Positron Emission Topography (PET) studies forexamining substrate receptor occupancy. Isotopically-labeled compoundsof structure (I) can generally be prepared by conventional techniquesknown to those skilled in the art or by processes analogous to thosedescribed in the Preparations and Examples as set out below using anappropriate isotopically-labeled reagent in place of the non-labeledreagent previously employed.

Isotopically-labeled compounds as disclosed herein can generally beprepared by conventional techniques known to those skilled in the art orby processes analogous to those described in the accompanying examplesand schemes using an appropriate isotopically-labeled reagent in placeof the non-labeled reagent previously employed.

Certain of the compounds as disclosed herein may exist as stereoisomers(i.e., isomers that differ only in the spatial arrangement of atoms)including optical isomers and conformational isomers (or conformers).The compounds disclosed herein include all stereoisomers, both as pureindividual stereoisomer preparations and enriched preparations of each,and both the racemic mixtures of such stereoisomers as well as theindividual diastereomers and enantiomers that may be separated accordingto methods that are known to those skilled in the art. Additionally, thecompounds disclosed herein include all tautomeric forms of thecompounds.

Certain of the compounds disclosed herein may exist as atropisomers,which are conformational stereoisomers that occur when rotation about asingle bond in the molecule is prevented, or greatly slowed, as a resultof steric interactions with other parts of the molecule. The compoundsdisclosed herein include all atropisomers, both as pure individualatropisomer preparations, enriched preparations of each, or anon-specific mixture of each. Where the rotational barrier about thesingle bond is high enough, and interconversion between conformations isslow enough, separation and isolation of the isomeric species may bepermitted.

EMBODIMENTS Embodiment 1

In one embodiment of the present invention, the present inventioncomprises a compound having formula (I):

-   -   wherein        -   R¹ is a —C₁-C₆ alkyl, or —C₃-C₆cycloalkyl, group;        -   R^(1a) is a —C₁-C₆ alkyl, —C₁-C₆ heteroalkyl, aryl,            heteroaryl, —C₃-C₆ cycloalkyl or —C₃-C₆ heterocycloalkyl            group;        -   R¹ and R^(1a) together with the carbon atom to which they            are attached, form a carbocyclic ring, wherein the            carbocyclic ring can be unsubstituted or fused to an            aromatic ring;        -   R² is an aryl substituted with a halo, —OH, or NH₂;        -   R³ is halo;        -   R⁴ is H or methyl;        -   R⁴ is H or methyl; or    -   a stereoisomer thereof, an atropisomer thereof, a        pharmaceutically acceptable salt thereof, a pharmaceutically        acceptable salt of the stereoisomer thereof, or a        pharmaceutically acceptable salt of the atropisomer thereof.

Embodiment 2

In another embodiment of the present invention, the present inventioncomprises a compound of embodiment 1 wherein R¹ is an —C₁-C₆ alkyl or—C₃-C₆ cycloalkyl group.

Embodiment 3

In another embodiment of the present invention, the present inventioncomprises a compound of embodiment 1 wherein R¹ is methyl.

Embodiment 4

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 1 wherein R¹ is a cyclopropylgroup.

Embodiment 5

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 1 wherein R^(1a) is an —C₁-C₆alkyl, aryl, or —C₃-C₆ cycloalkyl group.

Embodiment 6

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 5 wherein R^(1a) is an ethyl group.

Embodiment 7

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 5 wherein R^(1a) is a branched C₄alkyl group.

Embodiment 8

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 5 wherein R^(1a) is a cyclopropylgroup.

Embodiment 9

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 5 wherein R^(1a) is a cyclobutylgroup.

Embodiment 10

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 5 wherein R^(1a) is a cyclopentylgroup.

Embodiment 11

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 5 wherein R^(1a) is a phenyl group.

Embodiment 12

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 1 wherein R¹ and R^(1a), togetherwith the carbon atom to which they are attached, form a 4-10 memberedring.

Embodiment 13

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 12 wherein R¹ and R^(1a), togetherwith the carbon atom to which they are attached, form a cyclopentane.

Embodiment 14

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 13, wherein R¹ and R^(1a), togetherwith the carbon atom to which they are attached, form a cyclohexane.

Embodiment 15

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 12, wherein R¹ and R^(1a) togetherwith the carbon atom to which they are attached, form a 5-memberedcarbocyclic ring, wherein the carbocyclic ring can be unsubstituted orfused to an aromatic ring.

Embodiment 16

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 1, wherein R² is a fluorinatedphenyl.

Embodiment 17

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 1, wherein R³ is Cl.

Embodiment 18

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 1, wherein R⁴ is H.

Embodiment 19

In another embodiment of the present invention, the present inventioncomprises the compound of embodiment 1, wherein R⁴ is methyl.

Embodiment 20

In another embodiment of the present invention, the present inventioncomprises a compound having a structure selected from the formula

or a stereoisomer thereof, an atropisomer thereof, a pharmaceuticallyacceptable salt thereof, a pharmaceutically acceptable salt of thestereoisomer thereof, or a pharmaceutically acceptable salt of theatropisomer thereof.

Embodiment 21

In another embodiment of the present invention, the present inventioncomprises the compound of any one of embodiments 1-20 in the form of apharmaceutically acceptable salt.

Embodiment 22

In another embodiment of the present invention, the present inventioncomprises a pharmaceutical composition comprising the compound of anyone of embodiments 1-21 and a pharmaceutically acceptable excipient.

Embodiment 23

In another embodiment of the present invention, the present inventioncomprises a method of inhibiting KRAS G12C in a cell, comprisingcontacting the cell with the compound of any one of embodiments 1-21 orthe composition of embodiment 22.

Embodiment 24

In another embodiment of the present invention, the present inventioncomprises a method of treating cancer in a subject comprisingadministering to the subject a therapeutically effective amount of thecompound of any one of embodiments 1-21 or the composition of embodiment22.

Embodiment 25

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 24, wherein the cancer is lungcancer, pancreatic cancer, or colorectal cancer.

Embodiment 26

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 25, wherein the cancer is lungcancer.

Embodiment 27

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 25, wherein the cancer is pancreaticcancer.

Embodiment 28

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 25, wherein the cancer is colorectalcancer.

Embodiment 29

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 24, further comprising administeringto the patient in need thereof a therapeutically effective amount of oneor more additional pharmaceutically active compounds.

Embodiment 30

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 29, wherein the one or moreadditional pharmaceutically active compounds is an anti-PD-1 antibody.

Embodiment 31

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 29, wherein the one or moreadditional pharmaceutically active compounds is pembrolizumab.

Embodiment 32

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 29, wherein the one or moreadditional pharmaceutically active compounds is nivolumab.

Embodiment 33

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 29, wherein the one or moreadditional pharmaceutically active compounds is an MCI-1 inhibitor.

Embodiment 34

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 33, wherein the MCI-1 inhibitor isAMG-176.

Embodiment 35

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 29, wherein the one or moreadditional pharmaceutically active compounds is daratumumab.

Embodiment 36

In another embodiment of the present invention, the present inventioncomprises the method of embodiment 29, wherein the one or moreadditional pharmaceutically active compounds is an immunomodulatoryiMID.

Embodiment 37

The method of claim 29, wherein the one or more additionalpharmaceutically active compounds is a MEK inhibitor.

Embodiment 38

In another embodiment of the present invention, the present inventioncomprises a compound according to Claim 1 in the preparation of amedicament for treating cancer.

Embodiment 39

In another embodiment of the present invention, the present inventioncomprises the compound according to embodiment 26, wherein the cancer isnon-small cell lung cancer.

Synthesis of Disclosed Compounds

Compounds as disclosed herein can be synthesized via a number ofspecific methods. The examples which outline specific synthetic routes,and the generic schemes below are meant to provide guidance to theordinarily skilled synthetic chemist, who will readily appreciate thatthe solvent, concentration, reagent, protecting group, order ofsynthetic steps, time, temperature, and the like can be modified asnecessary, well within the skill and judgment of the ordinarily skilledartisan.

Pharmaceutical Compositions, Dosing, and Routes of Administration

Also provided herein are pharmaceutical compositions that include acompound as disclosed herein, together with a pharmaceuticallyacceptable excipient, such as, for example, a diluent or carrier.Compounds and pharmaceutical compositions suitable for use in thepresent invention include those wherein the compound can be administeredin an effective amount to achieve its intended purpose. Administrationof the compound described in more detail below.

Suitable pharmaceutical formulations can be determined by the skilledartisan depending on the route of administration and the desired dosage.See, e.g., Remington's Pharmaceutical Sciences, 1435-712 (18th ed., MackPublishing Co, Easton, Pa., 1990). Formulations may influence thephysical state, stability, rate of in vivo release and rate of in vivoclearance of the administered agents. Depending on the route ofadministration, a suitable dose may be calculated according to bodyweight, body surface areas or organ size. Further refinement of thecalculations necessary to determine the appropriate treatment dose isroutinely made by those of ordinary skill in the art without undueexperimentation, especially in light of the dosage information andassays disclosed herein as well as the pharmacokinetic data obtainablethrough animal or human clinical trials.

The phrases “pharmaceutically acceptable” or “pharmacologicallyacceptable” refer to molecular entities and compositions that do notproduce adverse, allergic, or other untoward reactions when administeredto an animal or a human. As used herein, “pharmaceutically acceptable”includes any and all solvents, dispersion media, coatings, antibacterialand antifungal agents, isotonic and absorption delaying agents and thelike. The use of such excipients for pharmaceutically active substancesis well known in the art. Except insofar as any conventional media oragent is incompatible with the therapeutic compositions, its use intherapeutic compositions is contemplated. Supplementary activeingredients also can be incorporated into the compositions. In exemplaryembodiments, the formulation may comprise corn syrup solids, high-oleicsafflower oil, coconut oil, soy oil, L-leucine, calcium phosphatetribasic, L-tyrosine, L-proline, L-lysine acetate, DATEM (anemulsifier), L-glutamine, L-valine, potassium phosphate dibasic,L-isoleucine, L-arginine, L-alanine, glycine, L-asparagine monohydrate,L-serine, potassium citrate, L-threonine, sodium citrate, magnesiumchloride, L-histidine, L-methionine, ascorbic acid, calcium carbonate,L-glutamic acid, L-cystine dihydrochloride, L-tryptophan, L-asparticacid, choline chloride, taurine, m-inositol, ferrous sulfate, ascorbylpalmitate, zinc sulfate, L-carnitine, alpha-tocopheryl acetate, sodiumchloride, niacinamide, mixed tocopherols, calcium pantothenate, cupricsulfate, thiamine chloride hydrochloride, vitamin A palmitate, manganesesulfate, riboflavin, pyridoxine hydrochloride, folic acid,beta-carotene, potassium iodide, phylloquinone, biotin, sodium selenate,chromium chloride, sodium molybdate, vitamin D3 and cyanocobalamin.

The compound can be present in a pharmaceutical composition as apharmaceutically acceptable salt. As used herein, “pharmaceuticallyacceptable salts” include, for example base addition salts and acidaddition salts.

Pharmaceutically acceptable base addition salts may be formed withmetals or amines, such as alkali and alkaline earth metals or organicamines. Pharmaceutically acceptable salts of compounds may also beprepared with a pharmaceutically acceptable cation. Suitablepharmaceutically acceptable cations are well known to those skilled inthe art and include alkaline, alkaline earth, ammonium and quaternaryammonium cations. Carbonates or hydrogen carbonates are also possible.Examples of metals used as cations are sodium, potassium, magnesium,ammonium, calcium, or ferric, and the like. Examples of suitable aminesinclude isopropylamine, trimethylamine, histidine,N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine.

Pharmaceutically acceptable acid addition salts include inorganic ororganic acid salts. Examples of suitable acid salts include thehydrochlorides, formates, acetates, citrates, salicylates, nitrates,phosphates. Other suitable pharmaceutically acceptable salts are wellknown to those skilled in the art and include, for example, formic,acetic, citric, oxalic, tartaric, or mandelic acids, hydrochloric acid,hydrobromic acid, sulfuric acid or phosphoric acid; with organiccarboxylic, sulfonic, sulfo or phospho acids or N-substituted sulfamicacids, for example acetic acid, trifluoroacetic acid (TFA), propionicacid, glycolic acid, succinic acid, maleic acid, hydroxymaleic acid,methylmaleic acid, fumaric acid, malic acid, tartaric acid, lactic acid,oxalic acid, gluconic acid, glucaric acid, glucuronic acid, citric acid,benzoic acid, cinnamic acid, mandelic acid, salicylic acid,4-aminosalicylic acid, 2-phenoxybenzoic acid, 2-acetoxybenzoic acid,embonic acid, nicotinic acid or isonicotinic acid; and with amino acids,such as the 20 alpha amino acids involved in the synthesis of proteinsin nature, for example glutamic acid or aspartic acid, and also withphenylacetic acid, methanesulfonic acid, ethanesulfonic acid,2-hydroxyethanesulfonic acid, ethane 1,2-disulfonic acid,benzenesulfonic acid, 4-methylbenzenesulfonic acid, naphthalene2-sulfonic acid, naphthalene 1,5-disulfonic acid, 2- or3-phosphoglycerate, glucose 6-phosphate, N-cyclohexylsulfamic acid (withthe formation of cyclamates), or with other acid organic compounds, suchas ascorbic acid.

Pharmaceutical compositions containing the compounds disclosed hereincan be manufactured in a conventional manner, e.g., by conventionalmixing, dissolving, granulating, dragee-making, levigating, emulsifying,encapsulating, entrapping, or lyophilizing processes. Proper formulationis dependent upon the route of administration chosen.

For oral administration, suitable compositions can be formulated readilyby combining a compound disclosed herein with pharmaceuticallyacceptable excipients such as carriers well known in the art. Suchexcipients and carriers enable the present compounds to be formulated astablets, pills, dragees, capsules, liquids, gels, syrups, slurries,suspensions and the like, for oral ingestion by a patient to be treated.Pharmaceutical preparations for oral use can be obtained by adding acompound as disclosed herein with a solid excipient, optionally grindinga resulting mixture, and processing the mixture of granules, afteradding suitable auxiliaries, if desired, to obtain tablets or drageecores. Suitable excipients include, for example, fillers and cellulosepreparations. If desired, disintegrating agents can be added.Pharmaceutically acceptable ingredients are well known for the varioustypes of formulation and may be for example binders (e.g., natural orsynthetic polymers), lubricants, surfactants, sweetening and flavoringagents, coating materials, preservatives, dyes, thickeners, adjuvants,antimicrobial agents, antioxidants and carriers for the variousformulation types.

When a therapeutically effective amount of a compound disclosed hereinis administered orally, the composition typically is in the form of asolid (e.g., tablet, capsule, pill, powder, or troche) or a liquidformulation (e.g., aqueous suspension, solution, elixir, or syrup).

When administered in tablet form, the composition can additionallycontain a functional solid and/or solid carrier, such as a gelatin or anadjuvant. The tablet, capsule, and powder can contain about 1 to about95% compound, and preferably from about 15 to about 90% compound.

When administered in liquid or suspension form, a functional liquidand/or a liquid carrier such as water, petroleum, or oils of animal orplant origin can be added. The liquid form of the composition canfurther contain physiological saline solution, sugar alcohol solutions,dextrose or other saccharide solutions, or glycols. When administered inliquid or suspension form, the composition can contain about 0.5 toabout 90% by weight of a compound disclosed herein, and preferably about1 to about 50% of a compound disclosed herein. In one embodimentcontemplated, the liquid carrier is non-aqueous or substantiallynon-aqueous. For administration in liquid form, the composition may besupplied as a rapidly-dissolving solid formulation for dissolution orsuspension immediately prior to administration.

When a therapeutically effective amount of a compound disclosed hereinis administered by intravenous, cutaneous, or subcutaneous injection,the composition is in the form of a pyrogen-free, parenterallyacceptable aqueous solution. The preparation of such parenterallyacceptable solutions, having due regard to pH, isotonicity, stability,and the like, is within the skill in the art. A preferred compositionfor intravenous, cutaneous, or subcutaneous injection typicallycontains, in addition to a compound disclosed herein, an isotonicvehicle. Such compositions may be prepared for administration assolutions of free base or pharmacologically acceptable salts in watersuitably mixed with a surfactant, such as hydroxypropylcellulose.Dispersions also can be prepared in glycerol, liquid polyethyleneglycols, and mixtures thereof and in oils. Under ordinary conditions ofstorage and use, these preparations can optionally contain apreservative to prevent the growth of microorganisms.

Injectable compositions can include sterile aqueous solutions,suspensions, or dispersions and sterile powders for the extemporaneouspreparation of sterile injectable solutions, suspensions, ordispersions. In all embodiments the form must be sterile and must befluid to the extent that easy syringability exists. It must be stableunder the conditions of manufacture and storage and must resist thecontaminating action of microorganisms, such as bacteria and fungi, byoptional inclusion of a preservative. The carrier can be a solvent ordispersion medium containing, for example, water, ethanol, polyol (e.g.,glycerol, propylene glycol, and liquid polyethylene glycol, and thelike), suitable mixtures thereof, and vegetable oils. In one embodimentcontemplated, the carrier is non-aqueous or substantially non-aqueous.The proper fluidity can be maintained, for example, by the use of acoating, such as lecithin, by the maintenance of the required particlesize of the compound in the embodiment of dispersion and by the use ofsurfactants. The prevention of the action of microorganisms can bebrought about by various antibacterial and antifungal agents, forexample, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, andthe like. In many embodiments, it will be preferable to include isotonicagents, for example, sugars or sodium chloride. Prolonged absorption ofthe injectable compositions can be brought about by the use in thecompositions of agents delaying absorption, for example, aluminummonostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the activecompounds in the required amount in the appropriate solvent with variousof the other ingredients enumerated above, as required, followed byfiltered sterilization. Generally, dispersions are prepared byincorporating the various sterilized active ingredients into a sterilevehicle which contains the basic dispersion medium and the requiredother ingredients from those enumerated above. In the embodiment ofsterile powders for the preparation of sterile injectable solutions, thepreferred methods of preparation are vacuum-drying and freeze-dryingtechniques which yield a powder of the active ingredient plus anyadditional desired ingredient from a previously sterile-filteredsolution thereof.

Slow release or sustained release formulations may also be prepared inorder to achieve a controlled release of the active compound in contactwith the body fluids in the GI tract, and to provide a substantiallyconstant and effective level of the active compound in the blood plasma.For example, release can be controlled by one or more of dissolution,diffusion, and ion-exchange. In addition, the slow release approach mayenhance absorption via saturable or limiting pathways within the GItract. For example, the compound may be embedded for this purpose in apolymer matrix of a biological degradable polymer, a water-solublepolymer or a mixture of both, and optionally suitable surfactants.Embedding can mean in this context the incorporation of micro-particlesin a matrix of polymers. Controlled release formulations are alsoobtained through encapsulation of dispersed micro-particles oremulsified micro-droplets via known dispersion or emulsion coatingtechnologies.

For administration by inhalation, compounds of the present invention areconveniently delivered in the form of an aerosol spray presentation frompressurized packs or a nebulizer, with the use of a suitable propellant.In the embodiment of a pressurized aerosol, the dosage unit can bedetermined by providing a valve to deliver a metered amount. Capsulesand cartridges of, e.g., gelatin, for use in an inhaler or insufflatorcan be formulated containing a powder mix of the compound and a suitablepowder base such as lactose or starch.

The compounds disclosed herein can be formulated for parenteraladministration by injection (e.g., by bolus injection or continuousinfusion). Formulations for injection can be presented in unit dosageform (e.g., in ampules or in multidose containers), with an addedpreservative. The compositions can take such forms as suspensions,solutions, or emulsions in oily or aqueous vehicles, and can containformulatory agents such as suspending, stabilizing, and/or dispersingagents.

Pharmaceutical formulations for parenteral administration includeaqueous solutions of the compounds in water-soluble form. Additionally,suspensions of the compounds can be prepared as appropriate oilyinjection suspensions. Suitable lipophilic solvents or vehicles includefatty oils or synthetic fatty acid esters. Aqueous injection suspensionscan contain substances which increase the viscosity of the suspension.Optionally, the suspension also can contain suitable stabilizers oragents that increase the solubility of the compounds and allow for thepreparation of highly concentrated solutions. Alternatively, a presentcomposition can be in powder form for constitution with a suitablevehicle (e.g., sterile pyrogen-free water) before use.

Compounds disclosed herein also can be formulated in rectalcompositions, such as suppositories or retention enemas (e.g.,containing conventional suppository bases). In addition to theformulations described previously, the compounds also can be formulatedas a depot preparation. Such long-acting formulations can beadministered by implantation (e.g., subcutaneously or intramuscularly)or by intramuscular injection. Thus, for example, the compounds can beformulated with suitable polymeric or hydrophobic materials (forexample, as an emulsion in an acceptable oil) or ion exchange resins, oras sparingly soluble derivatives, for example, as a sparingly solublesalt.

In particular, a compound disclosed herein can be administered orally,buccally, or sublingually in the form of tablets containing excipients,such as starch or lactose, or in capsules or ovules, either alone or inadmixture with excipients, or in the form of elixirs or suspensionscontaining flavoring or coloring agents. Such liquid preparations can beprepared with pharmaceutically acceptable additives, such as suspendingagents. A compound also can be injected parenterally, for example,intravenously, intramuscularly, subcutaneously, or intracoronarily. Forparenteral administration, the compound is best used in the form of asterile aqueous solution which can contain other substances, forexample, salts, or sugar alcohols, such as mannitol, or glucose, to makethe solution isotonic with blood.

For veterinary use, a compound disclosed herein is administered as asuitably acceptable formulation in accordance with normal veterinarypractice. The veterinarian can readily determine the dosing regimen androute of administration that is most appropriate for a particularanimal.

In some embodiments, all the necessary components for the treatment ofKRAS-related disorder using a compound as disclosed herein either aloneor in combination with another agent or intervention traditionally usedfor the treatment of such disease may be packaged into a kit.Specifically, the present invention provides a kit for use in thetherapeutic intervention of the disease comprising a packaged set ofmedicaments that include the compound disclosed herein as well asbuffers and other components for preparing deliverable forms of saidmedicaments, and/or devices for delivering such medicaments, and/or anyagents that are used in combination therapy with the compound disclosedherein, and/or instructions for the treatment of the disease packagedwith the medicaments. The instructions may be fixed in any tangiblemedium, such as printed paper, or a computer readable magnetic oroptical medium, or instructions to reference a remote computer datasource such as a world wide web page accessible via the internet.

A “therapeutically effective amount” means an amount effective to treator to prevent development of, or to alleviate the existing symptoms of,the subject being treated. Determination of the effective amounts iswell within the capability of those skilled in the art, especially inlight of the detailed disclosure provided herein. Generally, a“therapeutically effective dose” refers to that amount of the compoundthat results in achieving the desired effect. For example, in onepreferred embodiment, a therapeutically effective amount of a compounddisclosed herein decreases KRAS activity by at least 5%, compared tocontrol, at least 10%, at least 15%, at least 20%, at least 25%, atleast 30%, at least 35%, at least 40%, at least 45%, at least 50%, atleast 55%, at least 60%, at least 65%, at least 70%, at least 75%, atleast 80%, at least 85%, or at least 90%.

The amount of compound administered can be dependent on the subjectbeing treated, on the subject's age, health, sex, and weight, the kindof concurrent treatment (if any), severity of the affliction, the natureof the effect desired, the manner and frequency of treatment, and thejudgment of the prescribing physician. The frequency of dosing also canbe dependent on pharmacodynamic effects on arterial oxygen pressures.However, the most preferred dosage can be tailored to the individualsubject, as is understood and determinable by one of skill in the art,without undue experimentation. This typically involves adjustment of astandard dose (e.g., reduction of the dose if the patient has a low bodyweight).

While individual needs vary, determination of optimal ranges ofeffective amounts of the compound is within the skill of the art. Foradministration to a human in the curative or prophylactic treatment ofthe conditions and disorders identified herein, for example, typicaldosages of the compounds of the present invention can be about 0.05mg/kg/day to about 50 mg/kg/day, for example at least 0.05 mg/kg, atleast 0.08 mg/kg, at least 0.1 mg/kg, at least 0.2 mg/kg, at least 0.3mg/kg, at least 0.4 mg/kg, or at least 0.5 mg/kg, and preferably 50mg/kg or less, 40 mg/kg or less, 30 mg/kg or less, 20 mg/kg or less, or10 mg/kg or less, which can be about 2.5 mg/day (0.5 mg/kg×5 kg) toabout 5000 mg/day (50 mg/kg×100 kg), for example. For example, dosagesof the compounds can be about 0.1 mg/kg/day to about 50 mg/kg/day, about0.05 mg/kg/day to about 10 mg/kg/day, about 0.05 mg/kg/day to about 5mg/kg/day, about 0.05 mg/kg/day to about 3 mg/kg/day, about 0.07mg/kg/day to about 3 mg/kg/day, about 0.09 mg/kg/day to about 3mg/kg/day, about 0.05 mg/kg/day to about 0.1 mg/kg/day, about 0.1mg/kg/day to about 1 mg/kg/day, about 1 mg/kg/day to about 10 mg/kg/day,about 1 mg/kg/day to about 5 mg/kg/day, about 1 mg/kg/day to about 3mg/kg/day, about 3 mg/day to about 500 mg/day, about 5 mg/day to about250 mg/day, about 10 mg/day to about 100 mg/day, about 3 mg/day to about10 mg/day, or about 100 mg/day to about 250 mg/day. Such doses may beadministered in a single dose or it may be divided into multiple doses.

Methods of Using KRAS G12C Inhibitors

The present disclosure provides a method of inhibiting RAS-mediated cellsignaling comprising contacting a cell with an effective amount of oneor more compounds disclosed herein. Inhibition of RAS-mediated signaltransduction can be assessed and demonstrated by a wide variety of waysknown in the art. Non-limiting examples include a showing of (a) adecrease in GTPase activity of RAS; (b) a decrease in GTP bindingaffinity or an increase in GDP binding affinity; (c) an increase in Koff of GTP or a decrease in K off of GDP; (d) a decrease in the levelsof signaling transduction molecules downstream in the RAS pathway, suchas a decrease in pMEK, pERK, or pAKT levels; and/or (e) a decrease inbinding of RAS complex to downstream signaling molecules including butnot limited to Raf. Kits and commercially available assays can beutilized for determining one or more of the above.

The disclosure also provides methods of using the compounds orpharmaceutical compositions of the present disclosure to treat diseaseconditions, including but not limited to conditions implicated by G12CKRAS, HRAS or NRAS mutation (e.g., cancer).

In some embodiments, a method for treatment of cancer is provided, themethod comprising administering an effective amount of any of theforegoing pharmaceutical compositions comprising a compound as disclosedherein to a subject in need thereof. In some embodiments, the cancer ismediated by a KRAS, HRAS or NRAS G12C mutation. In various embodiments,the cancer is pancreatic cancer, colorectal cancer or lung cancer. Insome embodiments, the cancer is gall bladder cancer, thyroid cancer, andbile duct cancer.

In some embodiments the disclosure provides method of treating adisorder in a subject in need thereof, wherein the said method comprisesdetermining if the subject has a KRAS, HRAS or NRAS G12C mutation and ifthe subject is determined to have the KRAS, HRAS or NRAS G12C mutation,then administering to the subject a therapeutically effective dose of atleast one compound as disclosed herein or a pharmaceutically acceptablesalt thereof.

The disclosed compounds inhibit anchorage-independent cell growth andtherefore have the potential to inhibit tumor metastasis. Accordingly,another embodiment the disclosure provides a method for inhibiting tumormetastasis, the method comprising administering an effective amount acompound disclosed herein.

KRAS, HRAS or NRAS G12C mutations have also been identified inhematological malignancies (e.g., cancers that affect blood, bone marrowand/or lymph nodes). Accordingly, certain embodiments are directed toadministration of a disclosed compounds (e.g., in the form of apharmaceutical composition) to a patient in need of treatment of ahematological malignancy. Such malignancies include, but are not limitedto leukemias and lymphomas. For example, the presently disclosedcompounds can be used for treatment of diseases such as Acutelymphoblastic leukemia (ALL), Acute myelogenous leukemia (AML), Chroniclymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), Chronicmyelogenous leukemia (CML), Acute monocytic leukemia (AMoL) and/or otherleukemias. In other embodiments, the compounds are useful for treatmentof lymphomas such as all subtypes of Hodgkins lymphoma or non-Hodgkinslymphoma. In various embodiments, the compounds are useful for treatmentof plasma cell malignancies such as multiple myeloma, mantle celllymphoma, and Waldenstrom's macroglubunemia.

Determining whether a tumor or cancer comprises a G12C KRAS, HRAS orNRAS mutation can be undertaken by assessing the nucleotide sequenceencoding the KRAS, HRAS or NRAS protein, by assessing the amino acidsequence of the KRAS, HRAS or NRAS protein, or by assessing thecharacteristics of a putative KRAS, HRAS or NRAS mutant protein. Thesequence of wild-type human KRAS, HRAS or NRAS is known in the art,(e.g. Accession No. NP203524).

Methods for detecting a mutation in a KRAS, HRAS or NRAS nucleotidesequence are known by those of skill in the art. These methods include,but are not limited to, polymerase chain reaction-restriction fragmentlength polymorphism (PCR-RFLP) assays, polymerase chain reaction-singlestrand conformation polymorphism (PCR-SSCP) assays, real-time PCRassays, PCR sequencing, mutant allele-specific PCR amplification (MASA)assays, direct sequencing, primer extension reactions, electrophoresis,oligonucleotide ligation assays, hybridization assays, TaqMan assays,SNP genotyping assays, high resolution melting assays and microarrayanalyses. In some embodiments, samples are evaluated for G12C KRAS, HRASor NRAS mutations by real-time PCR. In real-time PCR, fluorescent probesspecific for the KRAS, HRAS or NRAS G12C mutation are used. When amutation is present, the probe binds and fluorescence is detected. Insome embodiments, the KRAS, HRAS or NRAS G12C mutation is identifiedusing a direct sequencing method of specific regions (e.g., exon 2and/or exon 3) in the KRAS, HRAS or NRAS gene. This technique willidentify all possible mutations in the region sequenced.

Methods for detecting a mutation in a KRAS, HRAS or NRAS protein areknown by those of skill in the art. These methods include, but are notlimited to, detection of a KRAS, HRAS or NRAS mutant using a bindingagent (e.g., an antibody) specific for the mutant protein, proteinelectrophoresis and Western blotting, and direct peptide sequencing.

Methods for determining whether a tumor or cancer comprises a G12C KRAS,HRAS or NRAS mutation can use a variety of samples. In some embodiments,the sample is taken from a subject having a tumor or cancer. In someembodiments, the sample is a fresh tumor/cancer sample. In someembodiments, the sample is a frozen tumor/cancer sample. In someembodiments, the sample is a formalin-fixed paraffin-embedded sample. Insome embodiments, the sample is a circulating tumor cell (CTC) sample.In some embodiments, the sample is processed to a cell lysate. In someembodiments, the sample is processed to DNA or RNA.

The disclosure also relates to a method of treating a hyperproliferativedisorder in a mammal that comprises administering to said mammal atherapeutically effective amount of a compound as disclosed herein, or apharmaceutically acceptable salt thereof. In some embodiments, saidmethod relates to the treatment of a subject who suffers from a cancersuch as acute myeloid leukemia, cancer in adolescents, adrenocorticalcarcinoma childhood, AIDS-related cancers (e.g. Lymphoma and Kaposi'sSarcoma), anal cancer, appendix cancer, astrocytomas, atypical teratoid,basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer,brain stem glioma, brain tumor, breast cancer, bronchial tumors, Burkittlymphoma, carcinoid tumor, atypical teratoid, embryonal tumors, germcell tumor, primary lymphoma, cervical cancer, childhood cancers,chordoma, cardiac tumors, chronic lymphocytic leukemia (CLL), chronicmyelogenous leukemia (CML), chronic myeloproliferative disorders, coloncancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma,extrahepatic ductal carcinoma in situ (DCIS), embryonal tumors, CNScancer, endometrial cancer, ependymoma, esophageal cancer,esthesioneuroblastoma, ewing sarcoma, extracranial germ cell tumor,extragonadal germ cell tumor, eye cancer, fibrous histiocytoma of bone,gall bladder cancer, gastric cancer, gastrointestinal carcinoid tumor,gastrointestinal stromal tumors (GIST), germ cell tumor, gestationaltrophoblastic tumor, hairy cell leukemia, head and neck cancer, heartcancer, liver cancer, Hodgkin lymphoma, hypopharyngeal cancer,intraocular melanoma, islet cell tumors, pancreatic neuroendocrinetumors, kidney cancer, laryngeal cancer, lip and oral cavity cancer,liver cancer, lobular carcinoma in situ (LCIS), lung cancer, lymphoma,metastatic squamous neck cancer with occult primary, midline tractcarcinoma, mouth cancer, multiple endocrine neoplasia syndromes,multiple myeloma/plasma cell neoplasm, mycosis fungoides,myelodysplastic syndromes, myelodysplastic/myeloproliferative neoplasms,multiple myeloma, merkel cell carcinoma, malignant mesothelioma,malignant fibrous histiocytoma of bone and osteosarcoma, nasal cavityand paranasal sinus cancer, nasopharyngeal cancer, neuroblastoma,non-hodgkin lymphoma, non-small cell lung cancer (NSCLC), oral cancer,lip and oral cavity cancer, oropharyngeal cancer, ovarian cancer,pancreatic cancer, papillomatosis, paraganglioma, paranasal sinus andnasal cavity cancer, parathyroid cancer, penile cancer, pharyngealcancer, pleuropulmonary blastoma, primary central nervous system (CNS)lymphoma, prostate cancer, rectal cancer, transitional cell cancer,retinoblastoma, rhabdomyosarcoma, salivary gland cancer, skin cancer,stomach (gastric) cancer, small cell lung cancer, small intestinecancer, soft tissue sarcoma, T-Cell lymphoma, testicular cancer, throatcancer, thymoma and thymic carcinoma, thyroid cancer, transitional cellcancer of the renal pelvis and ureter, trophoblastic tumor, unusualcancers of childhood, urethral cancer, uterine sarcoma, vaginal cancer,vulvar cancer, or viral-induced cancer. In some embodiments, said methodrelates to the treatment of a non-cancerous hyperproliferative disordersuch as benign hyperplasia of the skin (e. g., psoriasis), restenosis,or prostate (e. g., benign prostatic hypertrophy (BPH)).

In some embodiments, the methods for treatment are directed to treatinglung cancers, the methods comprise administering an effective amount ofany of the above described compound (or a pharmaceutical compositioncomprising the same) to a subject in need thereof. In certainembodiments the lung cancer is a non-small cell lung carcinoma (NSCLC),for example adenocarcinoma, squamous-cell lung carcinoma or large-celllung carcinoma. In some embodiments, the lung cancer is a small celllung carcinoma. Other lung cancers treatable with the disclosedcompounds include, but are not limited to, glandular tumors, carcinoidtumors and undifferentiated carcinomas.

The disclosure further provides methods of modulating a G12C MutantKRAS, HRAS or NRAS protein activity by contacting the protein with aneffective amount of a compound of the disclosure. Modulation can beinhibiting or activating protein activity. In some embodiments, thedisclosure provides methods of inhibiting protein activity by contactingthe G12C Mutant KRAS, HRAS or NRAS protein with an effective amount of acompound of the disclosure in solution. In some embodiments, thedisclosure provides methods of inhibiting the G12C Mutant KPAS, HRAS orNRAS protein activity by contacting a cell, tissue, or organ thatexpresses the protein of interest. In some embodiments, the disclosureprovides methods of inhibiting protein activity in subject including butnot limited to rodents and mammal (e.g., human) by administering intothe subject an effective amount of a compound of the disclosure. In someembodiments, the percentage modulation exceeds 25%, 30%, 40%, 50%, 60%,70%, 80%, or 90%. In some embodiments, the percentage of inhibitingexceeds 25%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

In some embodiments, the disclosure provides methods of inhibiting KRAS,HRAS or NRAS G12C activity in a cell by contacting said cell with anamount of a compound of the disclosure sufficient to inhibit theactivity of KRAS, HRAS or NRAS G12C in said cell. In some embodiments,the disclosure provides methods of inhibiting KRAS, HRAS or NRAS G12Cactivity in a tissue by contacting said tissue with an amount of acompound of the disclosure sufficient to inhibit the activity of KRAS,HRAS or NRAS G12C in said tissue. In some embodiments, the disclosureprovides methods of inhibiting KRAS, HRAS or NRAS G12C activity in anorganism by contacting said organism with an amount of a compound of thedisclosure sufficient to inhibit the activity of KRAS, HRAS or NRAS G12Cin said organism. In some embodiments, the disclosure provides methodsof inhibiting KRAS, HRAS or NRAS G12C activity in an animal bycontacting said animal with an amount of a compound of the disclosuresufficient to inhibit the activity of KRAS, HRAS or NRAS G12C in saidanimal. In some embodiments, the disclosure provides methods ofinhibiting KRAS, HRAS or NRAS G12C activity in a mammal by contactingsaid mammal with an amount of a compound of the disclosure sufficient toinhibit the activity of KRAS, HRAS or NRAS G12C in said mammal. In someembodiments, the disclosure provides methods of inhibiting KRAS, HRAS orNRAS G12C activity in a human by contacting said human with n amount ofa compound of the disclosure sufficient to inhibit the activity of KRAS,HRAS or NRAS G12C in said human. The present disclosure provides methodsof treating a disease mediated by KRAS, HRAS or NRAS G12C activity in asubject in need of such treatment.

Combination Therapy:

The present disclosure also provides methods for combination therapiesin which an agent known to modulate other pathways, or other componentsof the same pathway, or even overlapping sets of target enzymes are usedin combination with a compound of the present disclosure, or apharmaceutically acceptable salt thereof. In one aspect, such therapyincludes but is not limited to the combination of one or more compoundsof the disclosure with chemotherapeutic agents, therapeutic antibodies,and radiation treatment, to provide a synergistic or additivetherapeutic effect.

Many chemotherapeutics are presently known in the art and can be used incombination with the compounds of the disclosure. In some embodiments,the chemotherapeutic is selected from the group consisting of mitoticinhibitors, alkylating agents, anti-metabolites, intercalatingantibiotics, growth factor inhibitors, cell cycle inhibitors, enzymes,topoisomerase inhibitors, biological response modifiers, anti-hormones,angiogenesis inhibitors, and anti-androgens. Non-limiting examples arechemotherapeutic agents, cytotoxic agents, and non-peptide smallmolecules such as Gleevec® (Imatinib Mesylate), Kyprolis® (carfilzomib),Velcade® (bortezomib), Casodex (bicalutamide), Iressa®® (gefitinib),Venclexta™ (venetoclax) and Adriamycin™, (docorubicin) as well as a hostof chemotherapeutic agents. Non-limiting examples of chemotherapeuticagents include alkylating agents such as thiotepa and cyclosphosphamide(Cytoxan™); alkyl sulfonates such as busulfan, improsulfan andpiposulfan; aziridines such as benzodopa, carboquone, meturedopa, anduredopa; ethylenimines and methylamelamines including altretamine,triethylenemelamine, trietylenephosphoramide,triethylenethiophosphaoramide and trimethylololomelamine; nitrogenmustards such as chlorambucil, chlornaphazine, chlorocyclophosphamide,estramustine, ifosfamide, mechlorethamine, mechlorethamine oxidehydrochloride, melphalan, novembichin, phenesterine, prednimustine,trofosfamide, uracil mustard; nitrosureas such as carmustine,chlorozotocin, fotemustine, lomustine, nimustine, ranimustine;antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine,bleomycins, cactinomycin, calicheamicin, carabicin, carminomycin,carzinophilin, Casodex™, chromomycins, dactinomycin, daunorubicin,detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin,esorubicin, idarubicin, marcellomycin, mitomycins, mycophenolic acid,nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin,quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexateand 5-fluorouracil (5-FU); folic acid analogues such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine,androgens such as calusterone, dromostanolone propionate, epitiostanol,mepitiostane, testolactone; anti-adrenals such as aminoglutethimide,mitotane, trilostane; folic acid replenisher such as frolinic acid;aceglatone; aldophosphamide glycoside; aminolevulinic acid; amsacrine;bestrabucil; bisantrene; edatraxate; defofamine; demecolcine;diaziquone; elfomithine; elliptinium acetate; etoglucid; galliumnitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone;mopidamol; nitracrine; pentostatin; phenamet; pirarubicin; podophyllinicacid; 2-ethylhydrazide; procarbazine; PSK; razoxane; sizofiran;spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-trichlorotriethylamine; urethan; vindesine; dacarbazine;mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine;arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxanes, e.g.paclitaxel and docetaxel; retinoic acid; esperamicins; capecitabine; andpharmaceutically acceptable salts, acids or derivatives of any of theabove.

Also included as suitable chemotherapeutic cell conditioners areanti-hormonal agents that act to regulate or inhibit hormone action ontumors such as anti-estrogens including for example tamoxifen,(Nolvadex™), raloxifene, aromatase inhibiting 4(5)-imidazoles,4-hydroxytamoxifen, trioxifene, keoxifene, LY 117018, onapristone, andtoremifene (Fareston); and anti-androgens such as flutamide, nilutamide,bicalutamide, leuprolide, and goserelin; chlorambucil; gemcitabine;6-thioguanine; mercaptopurine; methotrexate; platinum analogs such ascisplatin and carboplatin; vinblastine; platinum; etoposide (VP-16);ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine;navelbine; novantrone; teniposide; daunomycin; aminopterin; xeloda;ibandronate, camptothecin-11 (CPT-11); topoisomerase inhibitor RFS 2000;difluoromethylornithine (DMFO).

Where desired, the compounds or pharmaceutical composition of thepresent disclosure can be used in combination with commonly prescribedanti-cancer drugs such as Herceptin®, Avastin®, Erbitux®, Rituxan®,Taxol®, Arimidex®, Taxotere®, ABVD, AVICINE, Abagovomab, Acridinecarboxamide, Adecatumumab, 17-N-Allylamino-17-demethoxygeldanamycin,Alpharadin, Alvocidib, 3-Aminopyridine-2-carboxaldehydethiosemicarbazone, Amonafide, Anthracenedione, Anti-CD22 immunotoxins,Antineoplastic, Antitumorigenic herbs, Apaziquone, Atiprimod,Azathioprine, Belotecan, Bendamustine, BIBW 2992, Biricodar,Brostallicin, Bryostatin, Buthionine sulfoximine, CBV (chemotherapy),Calyculin, cell-cycle nonspecific antineoplastic agents, Dichloroaceticacid, Discodermolide, Elsamitrucin, Enocitabine, Epothilone, Eribulin,Everolimus, Exatecan, Exisulind, Ferruginol, Forodesine, Fosfestrol, ICEchemotherapy regimen, IT-101, Imexon, Imiquimod, Indolocarbazole,Irofulven, Laniquidar, Larotaxel, Lenalidomide, Lucanthone, Lurtotecan,Mafosfamide, Mitozolomide, Nafoxidine, Nedaplatin, Olaparib, Ortataxel,PAC-1, Pawpaw, Pixantrone, Proteasome inhibitor, Rebeccamycin,Resiquimod, Rubitecan, SN-38, Salinosporamide A, Sapacitabine, StanfordV, Swainsonine, Talaporfin, Tariquidar, Tegafur-uracil, Temodar,Tesetaxel, Triplatin tetranitrate, Tris(2-chloroethyl)amine,Troxacitabine, Uramustine, Vadimezan, Vinflunine, ZD6126 or Zosuquidar.

This disclosure further relates to a method for using the compounds orpharmaceutical compositions provided herein, in combination withradiation therapy for inhibiting abnormal cell growth or treating thehyperproliferative disorder in the mammal. Techniques for administeringradiation therapy are known in the art, and these techniques can be usedin the combination therapy described herein. The administration of thecompound of the disclosure in this combination therapy can be determinedas described herein.

Radiation therapy can be administered through one of several methods, ora combination of methods, including without limitation external-beamtherapy, internal radiation therapy, implant radiation, stereotacticradiosurgery, systemic radiation therapy, radiotherapy and permanent ortemporary interstitial brachytherapy. The term “brachytherapy,” as usedherein, refers to radiation therapy delivered by a spatially confinedradioactive material inserted into the body at or near a tumor or otherproliferative tissue disease site. The term is intended withoutlimitation to include exposure to radioactive isotopes (e.g. At-211,I-131, I-125, Y-90, Re-186, Re-188, Sm-153, Bi-212, P-32, andradioactive isotopes of Lu). Suitable radiation sources for use as acell conditioner of the present disclosure include both solids andliquids. By way of non-limiting example, the radiation source can be aradionuclide, such as I-125, I-131, Yb-169, Ir-192 as a solid source,I-125 as a solid source, or other radionuclides that emit photons, betaparticles, gamma radiation, or other therapeutic rays. The radioactivematerial can also be a fluid made from any solution of radionuclide(s),e.g., a solution of I-125 or I-131, or a radioactive fluid can beproduced using a slurry of a suitable fluid containing small particlesof solid radionuclides, such as Au-198, Y-90. Moreover, theradionuclide(s) can be embodied in a gel or radioactive micro spheres.

The compounds or pharmaceutical compositions of the disclosure can beused in combination with an amount of one or more substances selectedfrom anti-angiogenesis agents, signal transduction inhibitors,antiproliferative agents, glycolysis inhibitors, or autophagyinhibitors.

Anti-angiogenesis agents, such as MMP-2 (matrix-metalloproteinase 2)inhibitors, MMP-9 (matrix-metalloproteinase 9) inhibitors, and COX-11(cyclooxygenase 11) inhibitors, can be used in conjunction with acompound of the disclosure and pharmaceutical compositions describedherein. Anti-angiogenesis agents include, for example, rapamycin,temsirolimus (CCI-779), everolimus (RAD001), sorafenib, sunitinib, andbevacizumab. Examples of useful COX-II inhibitors include alecoxib,valdecoxib, and rofecoxib. Examples of useful matrix metalloproteinaseinhibitors are described in WO 96/33172 WO 96/27583 European PatentPublication EP0818442, European Patent Publication EP1004578, WO98/07697, WO 98/03516, WO 98/34918, WO 98/34915, WO 98/33768, WO98/30566, European Patent Publication 606046, European PatentPublication 931 788, WO 90/05719, WO 99/52910, WO 99/52889, WO 99/29667,WO1999007675, European Patent Publication EP1786785, European PatentPublication No. EP1181017, United States Publication No. US20090012085,United States Publication U.S. Pat. No. 5,863,949, United StatesPublication U.S. Pat. No. 5,861,510, and European Patent PublicationEP0780386, all of which are incorporated herein in their entireties byreference. Preferred MMP-2 and MMP-9 inhibitors are those that havelittle or no activity inhibiting MMP-1. More preferred, are those thatselectively inhibit MMP-2 and/or AMP-9 relative to the othermatrix-metalloproteinases (i. e., MAP-1, MMP-3, MMP-4, MMP-5, MMP-6,MMP-7, MMP-8, MMP-10, MMP-11, MMP-12, and MMP-13). Some specificexamples of MMP inhibitors useful in the disclosure are AG-3340, RO32-3555, and RS 13-0830.

The present compounds may also be used in co-therapies with otheranti-neoplastic agents, such as acemannan, aclarubicin, aldesleukin,alemtuzumab, alitretinoin, altretamine, amifostine, aminolevulinic acid,amrubicin, amsacrine, anagrelide, anastrozole, ANCER, ancestim,ARGLABIN, arsenic trioxide, BAM 002 (Novelos), bexarotene, bicalutamide,broxuridine, capecitabine, celmoleukin, cetrorelix, cladribine,clotrimazole, cytarabine ocfosfate, DA 3030 (Dong-A), daclizumab,denileukin diftitox, deslorelin, dexrazoxane, dilazep, docetaxel,docosanol, doxercalciferol, doxifluridine, doxorubicin, bromocriptine,carmustine, cytarabine, fluorouracil, HIT diclofenac, interferon alfa,daunorubicin, doxorubicin, tretinoin, edelfosine, edrecolomab,eflornithine, emitefur, epirubicin, epoetin beta, etoposide phosphate,exemestane, exisulind, fadrozole, filgrastim, finasteride, fludarabinephosphate, formestane, fotemustine, gallium nitrate, gemcitabine,gemtuzumab zogamicin, gimeracil/oteracil/tegafur combination, glycopine,goserelin, heptaplatin, human chorionic gonadotropin, human fetal alphafetoprotein, ibandronic acid, idarubicin, (imiquimod, interferon alfa,interferon alfa, natural, interferon alfa-2, interferon alfa-2a,interferon alfa-2b, interferon alfa-N1, interferon alfa-_(n3),interferon alfacon-1, interferon alpha, natural, interferon beta,interferon beta-1a, interferon beta-1b, interferon gamma, naturalinterferon gamma-1a, interferon gamma-1b, interleukin-1 beta,iobenguane, irinotecan, irsogladine, lanreotide, LC 9018 (Yakult),leflunomide, lenograstim, lentinan sulfate, letrozole, leukocyte alphainterferon, leuprorelin, levamisole+fluorouracil, liarozole, lobaplatin,lonidamine, lovastatin, masoprocol, melarsoprol, metoclopramide,mifepristone, miltefosine, mirimostim, mismatched double stranded RNA,mitoguazone, mitolactol, mitoxantrone, molgramostim, nafarelin,naloxone+pentazocine, nartograstim, nedaplatin, nilutamide, noscapine,novel erythropoiesis stimulating protein, NSC 631570 octreotide,oprelvekin, osaterone, oxaliplatin, paclitaxel, pamidronic acid,pegaspargase, peginterferon alfa-2b, pentosan polysulfate sodium,pentostatin, picibanil, pirarubicin, rabbit antithymocyte polyclonalantibody, polyethylene glycol interferon alfa-2a, porfimer sodium,raloxifene, raltitrexed, rasburiembodiment, rhenium Re 186 etidronate,RII retinamide, rituximab, romurtide, samarium (153 Sm) lexidronam,sargramostim, sizofiran, sobuzoxane, sonermin, strontium-89 chloride,suramin, tasonermin, tazarotene, tegafur, temoporfin, temozolomide,teniposide, tetrachlorodecaoxide, thalidomide, thymalfasin, thyrotropinalfa, topotecan, toremifene, tositumomab-iodine 131, trastuzumab,treosulfan, tretinoin, trilostane, trimetrexate, triptorelin, tumornecrosis factor alpha, natural, ubenimex, bladder cancer vaccine,Maruyama vaccine, melanoma lysate vaccine, valrubicin, verteporfin,vinorelbine, VIRULIZIN, zinostatin stimalamer, or zoledronic acid;abarelix; AE 941 (Aeterna), ambamustine, antisense oligonucleotide,bcl-2 (Genta), APC 8015 (Dendreon), cetuximab, decitabine,dexaminoglutethimide, diaziquone, EL 532 (Elan), EM 800 (Endorecherche),eniluracil, etanidazole, fenretinide, filgrastim SD01 (Amgen),fulvestrant, galocitabine, gastrin 17 immunogen, HLA-B7 gene therapy(Vical), granulocyte macrophage colony stimulating factor, histaminedihydrochloride, ibritumomab tiuxetan, ilomastat, IM 862 (Cytran),interleukin-2, iproxifene, LDI 200 (Milkhaus), leridistim, lintuzumab,CA 125 MAb (Biomira), cancer MAb (Japan Pharmaceutical Development),HER-2 and Fe MAb (Medarex), idiotypic 105AD7 MAb (CRC Technology),idiotypic CEA MAb (Tilex), LYM-1-iodine 131 MAb (Techniclone),polymorphic epithelial mucin-yttrium 90 MAb (Antisoma), marimastat,menogaril, mitumomab, motexafin gadolinium, MX 6 (Galderma), nelarabine,nolatrexed, P 30 protein, pegvisomant, pemetrexed, porfiromycin,prinomastat, RL 0903 (Shire), rubitecan, satraplatin, sodiumphenylacetate, sparfosic acid, SRL 172 (SR Pharma), SU 5416 (SUGEN), TA077 (Tanabe), tetrathiomolybdate, thaliblastine, thrombopoietin tinethyl etiopurpurin, tirapazamine, cancer vaccine (Biomira), melanomavaccine (New York University), melanoma vaccine (Sloan KetteringInstitute), melanoma oncolysate vaccine (New York Medical College),viral melanoma cell lysates vaccine (Royal Newcastle Hospital), orvalspodar.

The compounds of the invention may further be used with VEGFRinhibitors. Other compounds described in the following patents andpatent applications can be used in combination therapy: U.S. Pat. No.6,258,812, US 2003/0105091, WO 01/37820, U.S. Pat. No. 6,235,764, WO01/32651, U.S. Pat. Nos. 6,630,500, 6,515,004, 6,713,485, 5,521,184,5,770,599, 5,747,498, WO 02/68406, WO 02/66470, WO 02/55501, WO04/05279, WO 04/07481, WO 04/07458, WO 04/09784, WO 02/59110, WO99/45009, WO 00/59509, WO 99/61422, U.S. Pat. No. 5,990,141, WO00/12089, and WO 00/02871.

In some embodiments, the combination comprises a composition of thepresent invention in combination with at least one anti-angiogenicagent. Agents are inclusive of, but not limited to, in vitrosynthetically prepared chemical compositions, antibodies, antigenbinding regions, radionuclides, and combinations and conjugates thereof.An agent can be an agonist, antagonist, allosteric modulator, toxin or,more generally, may act to inhibit or stimulate its target (e.g.,receptor or enzyme activation or inhibition), and thereby promote celldeath or arrest cell growth.

Exemplary anti-angiogenic agents include ERBITUX™ (IMC-C225), KDR(kinase domain receptor) inhibitory agents (e.g., antibodies and antigenbinding regions that specifically bind to the kinase domain receptor),anti-VEGF agents (e.g., antibodies or antigen binding regions thatspecifically bind VEGF, or soluble VEGF receptors or a ligand bindingregion thereof) such as AVASTIN™ or VEGF-TRAP™, and anti-VEGF receptoragents (e.g., antibodies or antigen binding regions that specificallybind thereto), EGFR inhibitory agents (e.g., antibodies or antigenbinding regions that specifically bind thereto) such as Vectibix(panitumumab), IRESSA™ (gefitinib), TARCEVA™ (erlotinib), anti-Ang1 andanti-Ang2 agents (e.g., antibodies or antigen binding regionsspecifically binding thereto or to their receptors, e.g., Tie2/Tek), andanti-Tie2 kinase inhibitory agents (e.g., antibodies or antigen bindingregions that specifically bind thereto). The pharmaceutical compositionsof the present invention can also include one or more agents (e.g.,antibodies, antigen binding regions, or soluble receptors) thatspecifically bind and inhibit the activity of growth factors, such asantagonists of hepatocyte growth factor (HGF, also known as ScatterFactor), and antibodies or antigen binding regions that specificallybind its receptor “c-met”.

Other anti-angiogenic agents include Campath, IL-8, B-FGF, Tekantagonists (Ceretti et al., U.S. Publication No. 2003/0162712; U.S.Pat. No. 6,413,932), anti-TWEAK agents (e.g., specifically bindingantibodies or antigen binding regions, or soluble TWEAK receptorantagonists; see, Wiley, U.S. Pat. No. 6,727,225), ADAM distintegrindomain to antagonize the binding of integrin to its ligands (Fanslow etal., U.S. Publication No. 2002/0042368), specifically binding anti-ephreceptor and/or anti-ephrin antibodies or antigen binding regions (U.S.Pat. Nos. 5,981,245; 5,728,813; 5,969,110; 6,596,852; 6,232,447;6,057,124 and patent family members thereof), and anti-PDGF-BBantagonists (e.g., specifically binding antibodies or antigen bindingregions) as well as antibodies or antigen binding regions specificallybinding to PDGF-BB ligands, and PDGFR kinase inhibitory agents (e.g.,antibodies or antigen binding regions that specifically bind thereto).

Additional anti-angiogenic/anti-tumor agents include: SD-7784 (Pfizer,USA); cilengitide. (Merck KGaA, Germany, EPO 770622); pegaptaniboctasodium, (Gilead Sciences, USA); Alphastatin, (BioActa, UK); M-PGA,(Celgene, USA, U.S. Pat. No. 5,712,291); ilomastat, (Arriva, USA, U.S.Pat. No. 5,892,112); emaxanib, (Pfizer, USA, U.S. Pat. No. 5,792,783);vatalanib, (Novartis, Switzerland); 2-methoxyestradiol, (EntreMed, USA);TLC ELL-12, (Elan, Ireland) anecortave acetate, (Alcon, USA); alpha-D148Mab, (Amgen, USA); CEP-7055, (Cephalon, USA); anti-Vn Mab, (Crucell,Netherlands) DAC:antiangiogenic, (ConjuChem, Canada); Angiocidin,(InKine Pharmaceutical, USA); KM-2550, (Kyowa Hakko, Japan); SU-0879,(Pfizer, USA); CGP-79787, (Novartis, Switzerland, EP 970070); ARGENTtechnology, (Ariad, USA); YIGSR-Stealth, (Johnson & Johnson, USA);fibrinogen-E fragment, (BioActa, UK); angiogenesis inhibitor, (Trigen,UK); TBC-1635, (Encysive Pharmaceuticals, USA); SC-236, (Pfizer, USA);ABT-567, (Abbott, USA); Metastatin, (EntreMed, USA); angiogenesisinhibitor, (Tripep, Sweden); maspin, (Sosei, Japan); 2-methoxyestradiol,(Oncology Sciences Corporation, USA); ER-68203-00, (IVAX, USA); Benefin,(Lane Labs, USA); Tz-93, (Tsumura, Japan); TAN-1120, (Takeda, Japan);FR-111142, (Fujisawa, Japan, JP 02233610); platelet factor 4, (RepliGen,USA, EP 407122); vascular endothelial growth factor antagonist, (Borean,Denmark); bevacizumab (pINN), (Genentech, USA); angiogenesis inhibitors,(SUGEN, USA); XL 784, (Exelixis, USA); XL 647, (Exelixis, USA); MAb,alpha5beta3 integrin, second generation, (Applied Molecular Evolution,USA and MedImmune, USA); gene therapy, retinopathy, (Oxford BioMedica,UK); enzastaurin hydrochloride (USAN), (Lilly, USA); CEP 7055,(Cephalon, USA and Sanofi-Synthelabo, France); BC 1, (Genoa Institute ofCancer Research, Italy); angiogenesis inhibitor, (Alchemia, Australia);VEGF antagonist, (Regeneron, USA); rBPI 21 and BPI-derivedantiangiogenic, (XOMA, USA); PI 88, (Progen, Australia); cilengitide(pINN), (Merck KGaA, German; Munich Technical University, Germany,Scripps Clinic and Research Foundation, USA); cetuximnab (INN),(Aventis, France); AVE 8062, (Ajinomoto, Japan); AS 1404, (CancerResearch Laboratory, New Zealand); SG 292, (Telios, USA); Endostatin,(Boston Childrens Hospital, USA); ATN 161, (Attenuon, USA); ANGIOSTATIN,(Boston Childrens Hospital, USA); 2-methoxyestradiol, (Boston ChildrensHospital, USA); ZD 6474, (AstraZeneca, UK); ZD 6126, (AngiogenePharmaceuticals, UK); PPI 2458, (Praecis, USA); AZD 9935, (AstraZeneca,UK); AZD 2171, (AstraZeneca, UK); vatalanib (pINN), (Novartis,Switzerland and Schering AG, Germany); tissue factor pathway inhibitors,(EntreMed, USA); pegaptanib (Pinn), (Gilead Sciences, USA);xanthorrhizol, (Yonsei University, South Korea); vaccine, gene-based,VEGF-2, (Scripps Clinic and Research Foundation, USA); SPV5.2,(Supratek, Canada); SDX 103, (University of California at San Diego,USA); PX 478, (ProlX, USA); METASTATIN, (EntreMed, USA); troponin I,(Harvard University, USA); SU 6668, (SUGEN, USA); OXI 4503, (OXiGENE,USA); o-guanidines, (Dimensional Pharmaceuticals, USA); motuporamine C,(British Columbia University, Canada); CDP 791, (Celltech Group, UK);atiprimod (pINN), (GlaxoSmithKline, UK); E 7820, (Eisai, Japan); CYC381, (Harvard University, USA); AE 941, (Aeterna, Canada); vaccine,angiogenesis, (EntreMed, USA); urokinase plasminogen activatorinhibitor, (Dendreon, USA); oglufanide (pINN), (Melmotte, USA);HIF-1alfa inhibitors, (Xenova, UK); CEP 5214, (Cephalon, USA); BAY RES2622, (Bayer, Germany); Angiocidin, (InKine, USA); A6, (Angstrom, USA);KR 31372, (Korea Research Institute of Chemical Technology, SouthKorea); GW 2286, (GlaxoSmithKline, UK); EHT 0101, (ExonHit, France); CP868596, (Pfizer, USA); CP 564959, (OSI, USA); CP 547632, (Pfizer, USA);786034, (GlaxoSmithKline, UK); KRN 633, (Kirin Brewery, Japan); drugdelivery system, intraocular, 2-methoxyestradiol, (EntreMed, USA);anginex, (Maastricht University, Netherlands, and Minnesota University,USA); ABT 510, (Abbott, USA); AAL 993, (Novartis, Switzerland); VEGI,(ProteomTech, USA); tumor necrosis factor-alpha inhibitors, (NationalInstitute on Aging, USA); SU 11248, (Pfizer, USA and SUGEN USA); ABT518, (Abbott, USA); YH16, (Yantai Rongchang, China); S-3APG, (BostonChildrens Hospital, USA and EntreMed, USA); MAb, KDR, (ImClone Systems,USA); MAb, alpha5 beta1, (Protein Design, USA); KDR kinase inhibitor,(Celltech Group, UK, and Johnson & Johnson, USA); GFB 116, (SouthFlorida University, USA and Yale University, USA); CS 706, (Sankyo,Japan); combretastatin A4 prodrug, (Arizona State University, USA);chondroitinase AC, (IBEX, Canada); BAY RES 2690, (Bayer, Germany); AGM1470, (Harvard University, USA, Takeda, Japan, and TAP, USA); AG 13925,(Agouron, USA); Tetrathiomolybdate, (University of Michigan, USA); GCS100, (Wayne State University, USA) CV 247, (Ivy Medical, UK); CKD 732,(Chong Kun Dang, South Korea); MAb, vascular endothelium growth factor,(Xenova, UK); irsogladine (INN), (Nippon Shinyaku, Japan); RG 13577,(Aventis, France); WX 360, (Wilex, Germany); squalamine (pINN),(Genaera, USA); RPI 4610, (Sirna, USA); cancer therapy, (Marinova,Australia); heparanase inhibitors, (InSight, Israel); KL 3106, (Kolon,South Korea); Honokiol, (Emory University, USA); ZK CDK, (Schering AG,Germany); ZK Angio, (Schering AG, Germany); ZK 229561, (Novartis,Switzerland, and Schering AG, Germany); XMP 300, (XOMA, USA); VGA 1102,(Taisho, Japan); VEGF receptor modulators, (Pharmacopeia, USA);VE-cadherin-2 antagonists, (ImClone Systems, USA); Vasostatin, (NationalInstitutes of Health, USA); vaccine, Flk-1, (ImClone Systems, USA); TZ93, (Tsumura, Japan); TumStatin, (Beth Israel Hospital, USA); truncatedsoluble FLT 1 (vascular endothelial growth factor receptor 1), (Merck &Co, USA); Tie-2 ligands, (Regeneron, USA); and, thrombospondin 1inhibitor, (Allegheny Health, Education and Research Foundation, USA).

Autophagy inhibitors include, but are not limited to chloroquine,3-methyladenine, hydroxychloroquine (Plaquenil™), bafilomycin Al,5-amino-4-imidazole carboxamide riboside (AICAR), okadaic acid,autophagy-suppressive algal toxins which inhibit protein phosphatases oftype 2A or type 1, analogues of cAMP, and drugs which elevate cAMPlevels such as adenosine, LY204002, N6-mercaptopurine riboside, andvinblastine. In addition, antisense or siRNA that inhibits expression ofproteins including but not limited to ATG5 (which are implicated inautophagy), may also be used.

Additional pharmaceutically active compounds/agents that can be used inthe treatment of cancers and that can be used in combination with one ormore compound of the present invention include: epoetin alfa;darbepoetin alfa; panitumumab; pegfilgrastim; palifermin; filgrastim;denosumab; ancestim; AMG 102; AMG 176; AMG 386; AMG 479; AMG 655; AMG745; AMG 951; and AMG 706, or a pharmaceutically acceptable saltthereof.

In certain embodiments, a composition provided herein is conjointlyadministered with a chemotherapeutic agent. Suitable chemotherapeuticagents may include, natural products such as vinca alkaloids (e.g.,vinblastine, vincristine, and vinorelbine), paclitaxel,epidipodophyllotoxins (e.g., etoposide and teniposide), antibiotics(e.g., dactinomycin (actinomycin D), daunorubicin, doxorubicin, andidarubicin), anthracyclines, mitoxantrone, bleomycins, plicamycin(mithramycin), mitomycin, enzymes (e.g., L-asparaginase whichsystemically metabolizes L-asparagine and deprives cells which do nothave the capacity to synthesize their own asparagine), antiplateletagents, antiproliferative/antimitotic alkylating agents such as nitrogenmustards (e.g., mechlorethamine, cyclophosphamide and analogs,melphalan, and chlorambucil), ethylenimines and methylmelamines (e.g.,hexaamethylmelaamine and thiotepa), CDK inhibitors (e.g., seliciclib,UCN-01, P1446A-05, PD-0332991, dinaciclib, P27-00, AT-7519, RGB286638,and SCH727965), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g.,carmustine (BCNU) and analogs, and streptozocin), trazenes-dacarbazinine(DTIC), antiproliferative/antimitotic antimetabolites such as folic acidanalogs (e.g., methotrexate), pyrimidine analogs (e.g., fluorouracil,floxuridine, and cytarabine), purine analogs and related inhibitors(e.g., mercaptopurine, thioguanine, pentostatin and2-chlorodeoxyadenosine), aromatase inhibitors (e.g., anastrozole,exemestane, and letrozole), and platinum coordination complexes (e.g.,cisplatin and carboplatin), procarbazine, hydroxyurea, mitotane,aminoglutethimide, histone deacetylase (HDAC) inhibitors (e.g.,trichostatin, sodium butyrate, apicidan, suberoyl anilide hydroamnicacid, vorinostat, LBH 589, romidepsin, ACY-1215, and panobinostat), mTorinhibitors (e.g., temsirolimus, everolimus ridaforolimus, andsirolimus), KSP(Eg5) inhibitors (e.g., Array 520), DNA binding agents(e.g., Zalypsis), PI3K delta inhibitor (e.g., GS-1101 and TGR-1202),PI3K delta and gamma inhibitor (e.g., CAL-130), multi-kinase inhibitor(e.g., TG02 and sorafenib), hormones (e.g., estrogen) and hormoneagonists such as leutinizing hormone releasing hormone (LHRH) agonists(e.g., goserelin, leuprolide and triptorelin), BAFF-neutralizingantibody (e.g., LY2127399), IKK inhibitors, p38MAPK inhibitors,anti-IL-6 (e.g., CNTO328), telomerase inhibitors (e.g., GRN 163L),aurora kinase inhibitors (e.g., MLN8237), cell surface monoclonalantibodies (e.g., anti-CD38 (HUMAX-CD38), anti-CSI (e.g., elotuzumab),HSP90 inhibitors (e.g., 17 AAG and 1KOS 953), PI3K/Akt inhibitors (e.g.,perifosine), Akt inhibitor (e.g., GSK-2141795), PKC inhibitors (e.g.,enzastaurin), FTIs (e.g., Zarnestra™), anti-CD138 (e.g., BT062), Torc1/2specific kinase inhibitor (e.g., LNK128), kinase inhibitor (e.g.,GS-1101), ER/UPR targeting agent (e.g., MKC-3946), cFMS inhibitor (e.g.,ARRY-382), JAK1/2 inhibitor (e.g., CYT387), PARP inhibitor (e.g.,olaparib and veliparib (ABT-888)), BCL-2 antagonist. Otherchemotherapeutic agents may include mechlorethamine, camptothecin,ifosfamide, tamoxifen, raloxifene, gemcitabine, navelbine, sorafenib, orany analog or derivative variant of the foregoing.

The compounds of the present invention may also be used in combinationwith radiation therapy, hormone therapy, surgery and immunotherapy,which therapies are well known to those skilled in the art.

In certain embodiments, a pharmaceutical composition provided herein isconjointly administered with a steroid. Suitable steroids may include,but are not limited to, 21-acetoxypregnenolone, alclometasone,algestone, amcinonide, beclomethasone, betamethasone, budesonide,chloroprednisone, clobetasol, clocortolone, cloprednol, corticosterone,cortisone, cortivazol, deflazacort, desonide, desoximetasone,dexamethasone, diflorasone, diflucortolone, difuprednate, enoxolone,fluazacort, flucloronide, flumethasone, flunisolide, fluocinoloneacetonide, fluocinonide, fluocortin butyl, fluocortolone,fluorometholone, fluperolone acetate, fluprednidene acetate,fluprednisolone, flurandrenolide, fluticasone propionate, formocortal,halcinonide, halobetasol propionate, halometasone, hydrocortisone,loteprednol etabonate, mazipredone, medrysone, meprednisone,methylprednisolone, mometasone furoate, paramethasone, prednicarbate,prednisolone, prednisolone 25-diethylaminoacetate, prednisolone sodiumphosphate, prednisone, prednival, prednylidene, rimexolone, tixocortol,triamcinolone, triamcinolone acetonide, triamcinolone benetonide,triamcinolone hexacetonide, and salts and/or derivatives thereof. In aparticular embodiment, the compounds of the present invention can alsobe used in combination with additional pharmaceutically active agentsthat treat nausea. Examples of agents that can be used to treat nauseainclude: dronabinol; granisetron; metoclopramide; ondansetron; andproclorperazine; or a pharmaceutically acceptable salt thereof.

The compounds of the present invention may also be used in combinationwith an additional pharmaceutically active compound that disrupts orinhibits RAS-RAF-ERK or PI3K-AKT-TOR signaling pathways. In other suchcombinations, the additional pharmaceutically active compound is a PD-1and PD-L1 antagonist. The compounds or pharmaceutical compositions ofthe disclosure can also be used in combination with an amount of one ormore substances selected from EGFR inhibitors, MEK inhibitors, PI3Kinhibitors, AKT inhibitors, TOR inhibitors, Mcl-1 inhibitors, BCL-2inhibitors, SHP2 inhibitors, proteasome inhibitors, and immunetherapies, including monoclonal antibodies, immunomodulatory imides(IMiDs), anti-PD-1, anti-PDL-1, anti-CTLA4, anti-LAG1, and anti-OX40agents, GITR agonists, CAR-T cells, and BiTEs.

EGFR inhibitors include, but are not limited to, small moleculeantagonists, antibody inhibitors, or specific antisense nucleotide orsiRNA. Useful antibody inhibitors of EGFR include cetuximab (Erbitux),panitumumab (Vectibix), zalutumumab, nimotuzumab, and matuzumab. Smallmolecule antagonists of EGFR include gefitinib, erlotinib (Tarceva), andmost recently, lapatinib (TykerB). See e.g., Yan L, et. al.,Pharmacogenetics and Pharmacogenomics In Oncology Therapeutic AntibodyDevelopment, BioTechniques 2005; 39(4): 565-8, and Paez J G, et. al.,EGFR Mutations In Lung Cancer Correlation With Clinical Response toGefitinib Therapy, Science 2004; 304(5676): 1497-500.

Non-limiting examples of small molecule EGFR inhibitors include any ofthe EGFR inhibitors described in the following patent publications, andall pharmaceutically acceptable salts and solvates of said EGFRinhibitors: European Patent Application EP 520722, published Dec. 30,1992; European Patent Application EP 566226, published Oct. 20, 1993;PCT International Publication WO 96/33980, published Oct. 31, 1996; U.S.Pat. No. 5,747,498, issued May 5, 1998; PCT International Publication WO96/30347, published Oct. 3, 1996; European Patent Application EP 787772,published Aug. 6, 1997; PCT International Publication WO 97/30034,published Aug. 21, 1997; PCT International Publication WO 97/30044,published Aug. 21, 1997; PCT International Publication WO 97/38994,published Oct. 23, 1997; PCT International Publication WO 97/49688,published Dec. 31, 1997; European Patent Application EP 837063,published Apr. 22, 1998; PCI International Publication WO 98/02434,published Jan. 22, 1998; PCT International Publication WO 97/38983,published Oct. 23, 1997; PCT International Publication WO 95/19774,published Jul. 27, 1995; PCT International Publication WO 95/19970,published Jul. 27, 1995; PCT International Publication WO 97/13771,published Apr. 17, 1997; PCT International Publication WO 98/02437,published Jan. 22, 1998; PCT International Publication WO 98/02438,published Jan. 22, 1998; PCT International Publication WO 97/32881,published Sep. 12, 1997; German Application DE 19629652, published Jan.29, 1998; PCT International Publication WO 98/33798, published Aug. 6,1998; PCT International Publication WO 97/32880, published Sep. 12,1997; PCT International Publication WO 97/32880 published Sep. 12, 1997;European Patent Application EP 682027, published Nov. 15, 1995; PCTInternational Publication WO 97/02266, published Jan. 23, 197; PCTInternational Publication WO 97/27199, published Jul. 31, 1997; PCTInternational Publication WO 98/07726, published Feb. 26, 1998; PCTInternational Publication WO 97/34895, published Sep. 25, 1997; PCTInternational Publication WO 96/31510′, published Oct. 10, 1996; PCTInternational Publication WO 98/14449, published Apr. 9, 1998; PCTInternational Publication WO 98/14450, published Apr. 9, 1998; PCTInternational Publication WO 98/14451, published Apr. 9, 1998; PCTInternational Publication WO 95/09847, published Apr. 13, 1995; PCTInternational Publication WO 97/19065, published May 29, 1997; PCTInternational Publication WO 98/17662, published Apr. 30, 1998; U.S.Pat. No. 5,789,427, issued Aug. 4, 1998; U.S. Pat. No. 5,650,415, issuedJul. 22, 1997; U.S. Pat. No. 5,656,643, issued Aug. 12, 1997; PCTInternational Publication WO 99/35146, published Jul. 15, 1999; PCTInternational Publication WO 99/35132, published Jul. 15, 1999; PCTInternational Publication WO 99/07701, published Feb. 18, 1999; and PCTInternational Publication WO 92/20642 published Nov. 26, 1992.Additional non-limiting examples of small molecule EGFR inhibitorsinclude any of the EGFR inhibitors described in Traxler, P., 1998, Exp.Opin. Ther. Patents 8(12):1599-1625.

Antibody-based EGFR inhibitors include any anti-EGFR antibody orantibody fragment that can partially or completely block EGFR activationby its natural ligand. Non-limiting examples of antibody-based EGFRinhibitors include those described in Modjtahedi, H., et al., 1993, Br.J. Cancer 67:247-253; Teramoto, T., et al., 1996, Cancer 77:639-645;Goldstein et al., 1995, Clin. Cancer Res. 1:1311-1318; Huang, S. M., etal., 1999, Cancer Res. 15:59(8):1935-40; and Yang, X., et al., 1999,Cancer Res. 59:1236-1243. Thus, the EGFR inhibitor can be monoclonalantibody Mab E7.6.3 (Yang, 1999 supra), or Mab C225 (ATCC Accession No.HB-8508), or an antibody or antibody fragment having the bindingspecificity thereof.

MEK inhibitors include, but are not limited to, CI-1040, AZD6244,PD318088, PD98059, PD334581, RDEA119, ARRY-142886, ARRY-438162, andPD-325901.

PI3K inhibitors include, but are not limited to, wortmannin,17-hydroxywortmannin analogs described in WO 06/044453,4-[2-(1H-Indazol-4-yl)-6-[[4-(methylsulfonyl)piperazin-1-yl]methyl]thieno[3,2-d]pyrimidin-4-yl]morpholine(also known as GDC 0941 and described in PCT Publication Nos. WO09/036,082 and WO 09/055,730),2-Methyl-2-[4-[3-methyl-2-oxo-8-(quinolin-3-yl)-2,3-dihydroimidazo[4,5-c]quinolin-1-yl]phenyl]propionitrile(also known as BEZ 235 or NVP-BEZ 235, and described in PCT PublicationNo. WO 06/122806),(S)-1-(4-((2-(2-aminopyrimidin-5-yl)-7-methyl-4-morpholinothieno[3,2-d]pyrimidin-6-yl)methyl)piperazin-1-yl)-2-hydroxypropan-1-one(described in PCT Publication No. WO 2008/070740), LY294002(2-(4-Morpholinyl)-8-phenyl-4H-1-benzopyran-4-one available from AxonMedchem), PI 103 hydrochloride(3-[4-(4-morpholinylpyrido-[3′,2′:4,5]furo[3,2-d]pyrimidin-2-yl]phenolhydrochloride available from Axon Medchem), PIK 75(N′-[(1E)-(6-bromoimidazo[1,2-a]pyridin-3-yl)methylene]-N,2-dimethyl-5-nitrobenzenesulfono-hydrazidehydrochloride available from Axon Medchem), PIK 90(N-(7,8-dimethoxy-2,3-dihydroimidazo[1,2-c]quinazolin-5-yl)-nicotinamideavailable from Axon Medchem), GDC-0941 bismesylate(2-(1H-Indazol-4-yl)-6-(4-methanesulfonyl-piperazin-1-ylmethyl)-4-morpholin-4-yl-thieno[3,2-d]pyrimidinebismesylate available from Axon Medchem), AS-252424(5-[1-[5-(4-Fluoro-2-hydroxy-phenyl)-furan-2-yl]-meth-(Z)-ylidene]-thiazolidine-2,4-dioneavailable from Axon Medchem), and TGX-221(7-Methyl-2-(4-morpholinyl)-9-[1-(phenylamino)ethyl]-4H-pyrido-[1,2-a]pyrimidin-4-oneavailable from Axon Medchem), XL-765, and XL-147. Other PI3K inhibitorsinclude demethoxyviridin, perifosine, CAL101, PX-866, BEZ235, SF1126,INK1117, IPI-145, BKM120, XL147, XL765, Palomid 529, GSK1059615,ZSTK474, PWT33597, IC87114, TG100-115, CAL263, PI-103, GNE-477,CUDC-907, and AEZS-136.

AKT inhibitors include, but are not limited to, Akt-1-1 (inhibits Akt1)(Barnett et al. (2005) Biochem. J., 385 (Pt. 2), 399-408); Akt-1-1,2(inhibits Ak1 and 2) (Barnett et al. (2005) Biochem. J 385 (Pt. 2),399-408); API-59CJ-Ome (e.g., Jin et al. (2004) Br. J Cancer 91,1808-12); 1-H-imidazo[4,5-c]pyridinyl compounds (e.g., WO05011700);indole-3-carbinol and derivatives thereof (e.g., U.S. Pat. No.6,656,963; Sarkar and Li (2004) J Nutr. 134(12 Suppl), 3493S-3498S);perifosine (e.g., interferes with Akt membrane localization;Dasmahapatra et al. (2004) Clin. Cancer Res. 10(15), 5242-52, 2004);phosphatidylinositol ether lipid analogues (e.g., Gills and Dennis(2004) Expert. Opin. Investig. Drugs 13, 787-97); and triciribine (TCNor API-2 or NCI identifier: NSC 154020; Yang et al. (2004) Cancer Res.64, 4394-9).

TOR inhibitors include, but are not limited to, AP-23573, CCI-779,everolimus, RAD-001, rapamycin, temsirolimus, ATP-competitiveTORC1/TORC2 inhibitors, including PI-103, PP242, PP30 and Torin 1. OtherTOR inhibitors in FKBP12 enhancer; rapamycins and derivatives thereofincluding: CCI-779 (temsirolimus), R. D001 (Everolimus; WO 9409010) andAP23573; rapalogs, e.g. as disclosed in WO 98/02441 and WO 01/14387,e.g. AP23573, AP23464, or AP23841; 40-(2-hydroxyethyl)rapamycin,40-[3-hydroxy(hydroxymethyl)methylpropanoate]-rapamycin (also calledCC1779), 40-epi-(tetrazolyt)-rapamycin (also called ABT578),32-deoxorapamycin, 16-pentynyloxy-32(S)-dihydrorapanycin, and otherderivatives disclosed in WO 05005434; derivatives disclosed in U.S. Pat.No. 5,258,389, WO 94/090101, WO 92/05179, U.S. Pat. No. 5,118,677, U.S.Pat. No. 5,118,678, U.S. Pat. Nos. 5,100,883, 5,151,413, 5,120,842, WO93/111130, WO 94/02136, WO 94/02485, WO 95/14023, WO 94/02136, WO95/16691, WO 96/41807, WO 96/41807 and U.S. Pat. No. 5,256,790;phosphorus-containing rapamycin derivatives (e.g., WO 05016252);4H-1-benzopyran-4-one derivatives (e.g., U.S. Provisional ApplicationNo. 60/528,340).

MCI-1 inhibitors include, but are not limited to, AMG-176, MIK665, andS63845 The myeloid cell leukemia-1 (MCL-1) protein is one of the keyanti-apoptotic members of the B-cell lymphoma-2 (BCL-2) protein family.Over-expression of MCL-1 has been closely related to tumor progressionas well as to resistance, not only to traditional chemotherapies butalso to targeted therapeutics including BCL-2 inhibitors such asABT-263.

SHP inhibitors include, but are not limited to, SHP099.

Proteasome inhibitors include, but are not limited to, Kyprolis®(carfilzomib), Velcade® (bortezomib), and oprozomib.

Immune therapies include, but are not limited to, anti-PD-1 agents,anti-PDL-1 agents, anti-CTLA-4 agents, anti-LAG1-4 agents, and anti-OX40agents.

Monoclonal antibodies include, but are not limited to, Darzalex®(daratumumab), Herceptin® (trastuzumab), Avastin® (bevacizumab),Rituxan® (rituximab), Lucentis® (ranibizumab), and Eylea® (aflibercept).

Immunomodulatory imide drugs (IMiDs) are a class of immunomodulatorydrugs (drugs that adjust immune responses) containing an imide group.The IMiD class includes thalidomide and its analogues (lenalidomide,pomalidomide, and apremilast).

Anti-PD-1 antibodies include, but are not limited to, pembrolizumab(Keytruda®) and nivoluimab (Opdivo®). Exemplary anti-PD-1 antibodies andmethods for their use are described by Goldberg et al., Blood110(1):186-192 (2007), Thompson et al., Clin. Cancer Res.13(6):1757-1761 (2007), and Korman et al., International Application No.PCT/JP2006/309606 (publication no. WO 2006/121168 A1), each of which areexpressly incorporated by reference herein, include: Yervoy™(ipilimumab) or Tremelimumab (to CTLA-4), galiximab (to B7.1),BMS-936558 (to PD-1), MK-3475 (to PD-1), AMP224 (to B7DC), BMS-936559(to B7-H1), MPDL3280A (to B7-H1), MEDI-570 (to ICOS), AMG 404, AMG557(to B7H2), MGA271 (to B7H3), IMP321 (to LAG-3), BMS-663513 (to CD137),PF-05082566 (to CD137), CDX-1127 (to CD27), anti-OX40 (Providence HealthServices), huMAbOX40L (to OX40L), Atacicept (to TACI), CP-870893 (toCD40), Lucatumumab (to CD40), Dacetuzumab (to CD40), Muromonab-CD3 (toCD3), Ipilumumab (to CTLA-4). Immune therapies also include geneticallyengineered I-cells (e.g., CAR-T cells) and bispecific antibodies (e.g.BiTEs).

In a particular embodiment, the compounds of the present invention areused in combination with an anti-PD-1 antibody.

The present disclosure further provides nucleic acid sequences encodingthe anti-PD-1 antibody (or an antigen binding portion thereof).

GITR agonists include, but are not limited to, GITR fusion proteins andanti-GITR antibodies (e.g., bivalent anti-GITR antibodies), such as, aGITR fusion protein described in U.S. Pat. No. 6,111,090box.c, EuropeanPatent No.: 090505B1, U.S. Pat. No. 8,586,023. PCT Publication Nos.: WO2010/003118 and 2011/090754, or an anti-GITR antibody described, e.g.,in U.S. Pat. No. 7,025,962, European Patent No.: 1947183B1, U.S. Pat.Nos. 7,812,135, 8,388,967, 8,591,886, European Patent No.: EP 1866339,PCT Publication No.: WO 2011/028683, PCT Publication No.: WO2013/039954, PCT Publication No.: WO02005/007190, PCT Publication No.:WO 2007/133822, PCT Publication No.: WO2005/055808, PCT Publication No.:WO 99/40196, PCT Publication No.: WO 2001/03720, PCT Publication No.:WO99/20758, PCT Publication No.: WO2006/083289, PCT Publication No.: WO2005/115451, U.S. Pat. No. 7,618,632, and PCT Publication No.: WO2011/051726.

The compounds described herein can be used in combination with theagents disclosed herein or other suitable agents, depending on thecondition being treated. Hence, in some embodiments the one or morecompounds of the disclosure will be co-administered with other agents asdescribed above. When used in combination therapy, the compoundsdescribed herein are administered with the second agent simultaneouslyor separately. This administration in combination can includesimultaneous administration of the two agents in the same dosage form,simultaneous administration in separate dosage forms, and separateadministration. That is, a compound described herein and any of theagents described above can be formulated together in the same dosageform and administered simultaneously. Alternatively, a compound of thedisclosure and any of the agents described above can be simultaneouslyadministered, wherein both the agents are present in separateformulations. In another alternative, a compound of the presentdisclosure can be administered just followed by and any of the agentsdescribed above, or vice versa. In some embodiments of the separateadministration protocol, a compound of the disclosure and any of theagents described above are administered a few minutes apart, or a fewhours apart, or a few days apart.

As one aspect of the present invention contemplates the treatment of thedisease/conditions with a combination of pharmaceutically activecompounds that may be administered separately, the invention furtherrelates to combining separate pharmaceutical compositions in kit form.The kit comprises two separate pharmaceutical compositions: a compoundof the present invention, and a second pharmaceutical compound. The kitcomprises a container for containing the separate compositions such as adivided bottle or a divided foil packet. Additional examples ofcontainers include syringes, boxes, and bags. In some embodiments, thekit comprises directions for the use of the separate components. The kitform is particularly advantageous when the separate components arepreferably administered in different dosage forms (e.g., oral andparenteral), are administered at different dosage intervals, or whentitration of the individual components of the combination is desired bythe prescribing health care professional.

EXAMPLES Method 1 Example 1-15-((2S,5R)-4-Acryloyl-2,5-dimethylpiperazin-1-yl)-3-chloro-8-cyclopentyl-2-(2-fluorophenyl)-8-methyl-1,6-naphthyridin-7(8H)-one

Step 1.3-Chloro-8-cyclopentyl-2-(2-fluorophenyl)-8-methyl-1,6-naphthyridine-5,7(6H,8H)-dione

To a 100-mL round-bottomed flask was added 2-cyclopentylpropanenitrile(Intermediate I-1x, 0.691 g, 5.61 mmol) in toluene (3.51 mL). Thereaction mixture was cooled to 0° C. with an ice bath, then potassiumbis(trimethylsilyl)amide (1.0M in THF) (6.31 mL, 6.31 mmol,Sigma-Aldrich, St. Louis, Mo., USA) was added to the reaction mixture.The reaction mixture was allowed to stir at 0° C. while under an inert(N₂) atmosphere for 30 min. Then a heterogeneous mixture of2,5-dichloro-6-(2-fluorophenyl)nicotinamide (Intermediate 99B, 0.400 g1.4040 mol) in toluene (3.51 mL) was added to the reaction mixture. Theresulting reaction mixture was allowed to stir 30 min, while thetemperature was maintained at 0° C., then the ice bath was removed andthe reaction mixture was allowed to warm to ambient temperature over 2h. The reaction mixture was diluted with sat. aq. NaHCO₃ and EtOAc. Thelayers were separated and the aqueous layer was extracted with EtOAc.The combined organic extracts were dried over MgSO₄, filtered andconcentrated in vacuo.

The crude residue was treated with cone, hydrochloric acid, 37% a.c.s.reagent (2.60 mL, 84 mmol) and the reaction mixture was heated andstirred at 70° C. for 16 h. The mixture was removed from the heat bathand allowed to cool to ambient temperature. The mixture was made basicby slowly adding to a cold mixture of 10N NaOH with wet ice. Then CHCl₃was added to the mixture and the layers were separated. The aqueouslayer was extracted with CHCl₃. The combined organic extracts were driedover MgSO₄, filtered and concentrated in vacuo. The crude material wasabsorbed onto a plug of silica gel and purified by chromatographythrough a Redi-Sep pre-packed silica gel column (40 g), eluting with agradient of 0-5% MeOH in CH₂Cl₂, to provide3-chloro-8-cyclopentyl-2-(2-fluorophenyl)-8-methyl-1,6-naphthyridine-5,7(6H,8H)-dione(0.040 g, 0.10 mmol, 7.65% yield) as light-yellow solid. 1H NMR (400MHz, DMSO-d₆) δ 11.71 (s, 1H) 8.49 (s, 1H) 7.49-7.64 (m, 2H) 7.34-7.45(m, 2H) 2.34-2.42 (m, 1H) 1.62 (s, 4H) 1.33-1.44 (m, 7H). m/z (ESI, +veion): 373.1 (M+H)⁺.

Step 2.5-((2S,5R)-4-Acryloyl-2,5-dimethylpiperazin-1-yl)-3-chloro-8-cyclopentyl-2-(2-fluorophenyl)-8-methyl-1,6-naphthyridin-7(8H)-one

To a 50-mL round-bottomed flask was added3-chloro-8-cyclopentyl-2-(2-fluorophenyl)-8-methyl-1,6-naphthyridine-5,7(6H,8H)-dione(0.030 g, 0.08 mmol) in toluene (0.40 mL). ThenN,N′-diisopropylethylamine (0.14 mL, 0.80 mmol) and phosphorousoxychloride (0.03 mL, 0.40 mmol, Sigma-Aldrich, St. Louis, Mo., USA) wasadded to the reaction mixture. The resulting reaction mixture was heatedand stirred at 95° C. for 3 h, while under an inert (N₂) atmosphere. Theflask was removed from the heat bath and the mixture was allowed to coolto ambient temperature, then set aside.

To a 50-mL round-bottomed flask was added1-((2R,5S)-2,5-dimethylpiperazin-1-yl)prop-2-en-1-one2,2,2-trifluoroacetate (Intermediate 160, 0.06 g, 0.24 mmol) in EtOAc (1mL). Then DIPEA (1 mL) was added to the mixture, while stirred. To thismixture, was added the previous crude mixture in small portions. Afterthe addition, the reaction mixture was quenched with sat. aq. NH₄Cl,then the reaction mixture was diluted with EtOAc. The layers wereseparated and the aqueous layer was extracted with EtOAc. The combinedorganic extracts were dried over MgSO₄, filtered and concentrated invacuo. The crude material was absorbed onto a plug of silica, gel andpurified by chromatography through an Interchim (15 micron) silica-gelcolumn (25 grams), eluting with a gradient of 0-80% EtOAc in DCM, toprovide5-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-3-chloro-8-cyclopentyl-2-(2-fluorophenyl)-8-methyl-1,6-naphthyridin-7(8H)-one(0.015 g, 0.02 mmol, 35.6% yield) as a white solid diastereomericmixture. ¹H NMR (400 MHz, DMSO-d₆) δ 8.21 (br s, 1H) 7.46-7.57 (m, 2H)7.28-7.36 (m, 2H) 6.65-6.83 (m, 1H) 6.11 (br d, J=17.21 Hz, 1H) 5.68 (brd, J=9.95 Hz, 1H) 4.73-5.11 (m, 1H) 4.64 (br s, 1H) 4.31-4.51 (m, 1H)3.75-3.96 (m, 1H) 3.43 (br d, J=12.23 Hz, 1H) 2.14 (br s, 1H) 1.42 (brs, 3H) 1.24-1.38 (m, 10H) 1.18 (br d, J=6.84 Hz, 3H) 1.03-1.13 (m, 1H)0.98 (br s, 1H). m/z (ESI, +ve ion): 523.1 (M+H)⁺.

TABLE 1 Compounds 1-2-1-8 were prepared following the proceduredescribed in Method 1, steps 1-2, above as follows: Method Ex. #Chemical Structure Name Changes Reagents 1-2

5-((2S,5R)-4-Acryloyl- 2,5-dimethylpiperazin- 1-yl)-3-chloro-8-cyclopropyl-2-(2- fluorophenyl)-8-methyl- 1,6-naphthyridin-7(8H)- oneStep 1. Use Intermedaite 1- 2x 1-3

5-((2S,5R)-4-Acryloyl- 2,5-dimethylpiperazin- 1-yl)-3-chloro-8-cyclobutyl-2-(2- fluorophenyl)-8-methyl- 1,6-naphthyridin-7(8H)- oneStep 1. Use intermediate 1- 3x 1-4

5-((2S,5R)-4-Acryloyl- 2,5-dimethylpiperazin- 1-yl)-3-chloro-8-ethyl-2-(2-fluorophenyl)-8- methyl-1,6- naphthyridin-7(8H)-one Step 1. 2-Methylbutane nitrile (Enamine Ltd., Monmouth, NJ, USA) 1-5

5-((2S,5R)-4-Acryloyl- 2,5-dimethylpiperazin- 1-yl)-3-chloro-2-(2-fluorophenyl)-8-methyl- 8-phenyl-1,6- naphthyridin-7(8H)-one Step 1.Alpha- methylbenzyl cyanide (Sigma- Aldrich, St. Louis, MO, USA) 1-6

5′-((2S,5R)-4-Acryloyl- 2,5-dimethylpiperazin- 1-yl)-3′-chloro-2′-(2-fluorophenyl)-7′H- spiro[cyclohexane-1,8′- [16,]naphthyridin]-7′- oneStep 1. Cyclohexane- carbonitrile (Combi- Blocks, San Diego, CA, USA)1-7

5′-((2S,5R)-4-Acryloyl- 2,5-dimethylpiperazin- 1-yl)-3′-chloro-2′-(2-fluorophenyl)-2,3- dihdyro-7′H- spiro[indene-1,8′-[1,6]naphthyridin]-7′- one Step 1. 2,3- Dihydro-1H- indene-1-carbonitrile (Enamine Ltd., Monmouth, NJ, USA) 1-8

5-((2S,5R)-4-Acryloyl- 2,5-dimethylpiperazin- 1-yl)-3-chloro-2-(2-fluorophenyl)-8- isobutyl-8-methyl-1,6- naphthyridin-7(8H)-one Step 1.Use Intermediate 1- 4x

Method 2 Example 2-15-((2S,5R)-4-Acryloyl-2,5-dimethylpiperazin-1-yl)-3-chloro-8,8-dicyclopropyl-2-(2-fluorophenyl)-1,6-naphthyridin-7(8H)-one

Step 1.3-Chloro-8,8-dicyclopropyl-2-(2-fluorophenyl)-1,6-naphthyridine-5,7(6H,8H)-dione

To a 150-mL round-bottomed flask was added 2,2-dicyclopropylacetonitrile(Intermediate I-5x, 0.357 g, 2.95 mmol) in toluene (5.0 mL). Thereaction mixture was cooled to 0° C. in an ice bath, then potassiumbis(trimethylsilyl)amide (1.0M in THF) (3.31 mL, 3.31 mmol) was added tothe reaction mixture. The reaction mixture was allowed to stir at 0° C.while under an inert (N₂) atmosphere for 30 min. Then a solution of2,5-dichloro-6-(2-fluorophenyl)nicotinamide (Intermediate 99B, 0.210 g,0.73 mmol) in THF (3 mL) was added to the reaction mixture. Theresulting reaction mixture was allowed to stir 30 min, while thetemperature was maintained at 0° C., then the ice bath was removed andthe reaction mixture was allowed to warm to ambient temperature over 30min. The mixture was washed with sat. NaHCO₃, then the aqueous layer wasextracted with EtOAc. The combined organic extracts were dried overNa₂SO₄, filtered and concentrated in vacuo to afford5-chloro-2-(cyanodicyclopropylmethyl)-6-(2-fluorophenyl)nicotinamide.m/z (ESI, +ve ion): 370 (M+H)⁺.

The crude residue was treated with cone, hydrochloric acid, 37% a.c.s.reagent (6.0 mL, 194 mmol) and the reaction mixture was heated andstirred at 70° C. for 2 h. The reaction mixture was allowed to cool tort, then poured into ice, followed by the careful addition of cold 5NNaOH to basify it. The mixture was extracted with DCM and the combinedorganics were purified by silica-gel chromatography, eluting with agradient of 0-50% EtOAc in heptane to afford3-chloro-8,8-dicyclopropyl-2-(2-fluorophenyl)-1,6-naphthyridine-5,7(6H,8H)-dione(0.130 g, 0.35 mmol, 47.6% yield) as an off-white solid. ¹H NMR (400MHz, DMSO-d₆) δ 11.72 (br s, 1H), 8.51 (s, 1H), 7.53-7.69 (m, 2H),7.37-7.47 (m, 2H), 1.49-1.59 (m, 2H), 0.51 (br d, J=5.2 Hz, 4H),0.34-0.46 (m, 4H). m/z (ESI, +ve ion): 370.6 (M+H)⁺.

Step 2. tert-Butyl(2R,5S)-4-(3-chloro-8,8-dicyclopropyl-2-(2-fluorophenyl)-7-oxo-7,8-dihydro-1,6-naphthyridin-5-yl)-2,5-dimethylpiperazine-1-carboxylate

To a mixture of3-chloro-8,8-dicyclopropyl-2-(2-fluorophenyl)-1,6-naphthyridine-5,7(6H,8H)-dione(0.130 g, 0.35 mmol) in toluene (5.0 mL) was addedN,N′-diisopropylethylamine (0.61 mL, 3.51 mmol). Then phosphorousoxychloride (0.16 mL, 1.75 mmol) was added to the reaction mixture. Theresulting mixture mixture was stirred and heated at 95° C. for 4 h. Thereaction mixture was brought to rt, then the mixture was cooled to 0° C.with a wet ice bath. Then DIPEA (10 eq) and tert-butyl(2R,5S)-2,5-dimethylpiperazine-1-carboxylate 2,2,2-trifluoroacetate(0.345 g, 1.05 mmol, Asta Tech, Bristol, Pa., USA) was added to thereaction mixture. The reaction mixture was stirred at 0° C. for 15 min,then the reaction mixture was allowed to warm to rt over 15 min. Thereaction mixture was washed with cold sat. aq. NaHCO₃ and the layerswere separated. The aqueous layer was extracted with EtOAc. The combinedorganic extracts were dried over Na₂SO₄, filtered and concentrated invacuo. The crude material was purified by silica gel chromatography,eluting with a gradient of 0-50% EtOAc in heptane to afford tert-butyl(2R,5S)-4-(3-chloro-8,8-dicyclopropyl-2-(2-fluorophenyl)-7-oxo-7,8-dihydro-1,6-naphthyridin-5-yl)-2,5-dimethylpiperazine-1-carboxylate(0.162 g, 0.28 mmol, 81% yield) as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ 8.12 (s, 1H), 7.55-7.65 (m, 2H), 7.36-7.44 (m, 2H), 4.77-4.95(m, 1H), 4.24-4.47 (m, 1H), 3.68-3.79 (m, 1H), 1.43-1.49 (m, 9H),1.30-1.36 (m, 4H), 1.25-1.28 (min, 4H), 1.16 (br d, J=6.4 Hz, 3H),0.35-0.53 (m, 8H). m/z (ESI, +ve ion): 566.6 (M+H)⁺.

Step 3.5-((2S,5R)-4-Acryloyl-2,5-dimethylpiperazin-1-yl)-3-chloro-8,8-dicyclopropyl-2-(2-fluorophenyl)-1,6-naphthyridin-7(8H)-one

To a solution of tert-butyl(2R,5S)-4-(3-chloro-8,8-dicyclopropyl-2-(2-fluorophenyl)-7-oxo-7,8-dihydro-1,6-naphthyridin-5-yl)-2,5-dimethylpiperazine-1-carboxylate(0.162 g, 0.28 mmol) in DCM (2 mL) was added TFA (4.0 mL, 51.9 mmol).The resulting reaction mixture was stirred at rt for 20 min, then themixture was concentrated in vacuo. The crude residue was carried intothe next step of the synthesis without further purification.

3-Chloro-8,8-dicyclopropyl-5-((2S,5R)-2,5-dimethylpiperazin-1-yl)-2-(2-fluorophenyl)-1,6-naphthyridin-7(8H)-onewas dissolved in DCM (5 mL). Then N,N-diisopropylethylamine (0.24 mL,1.42 mmol) and acryloyl chloride (0.02 mL, 0.31 mmol) was added to thereaction mixture. The resulting reaction mixture was stirred at rt for30 min. The reaction mixture was washed with sat. aq. NH₄Cl, thenextracted the aqueous layer with DCM. The combined organic extracts weredried over Na₂SO₄, filtered and concentrated in vacuo. The crudematerial was purified by silica-gel chromatography, eluting with agradient of 0-5% MeOH in DCM to afford5-((2S,5R)-4-acryloyl-2,5-dimethylpiperazin-1-yl)-3-chloro-8,8-dicyclopropyl-2-(2-fluorophenyl)-1,6-naphthyridin-7(8H)-one(0.112 g, 0.21 mmol, 75% yield) as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ 8.08-8.20 (m, 1H), 7.54-7.67 (m, 2H), 7.32-7.47 (m, 2H),6.74-6.93 (m, 1H), 6.12-6.24 (m, 1H), 5.75 (br d, J=10.0 Hz, 1H),4.74-5.03 (m, 1H), 4.39-4.59 (m, 1H), 4.12-4.30 (m, 1H), 3.82-3.95 (m,1H), 3.56-3.76 (m, 1H), 1.29-1.41 (m, 5H), 1.22-1.28 (m, 2H), 1.15-1.20(m, 2H), 0.32-0.60 (m, 8H). m/z (ESI, +ve ion): 520.6 (M+H)⁺.

TABLE 2 Compound 2-2 was prepared following the procedure described inMethod 2, steps 1-3, above as follows: Method Ex. # Chemical StructureName Changes Reagents 2-2

5′-((2S,5R)-4- acryloyl-2,5- dimethylpiperazin- 1-yl)-3′-chloro- 2′-(2-fluorophenyl)- 1,3-dihydro-7′H- spiro[indene-2,8′- [1,6]naphthyridin]-7′-one Step 1. Use intermediate 1-6x.

Method 3 Example 3-15-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-3-chloro-2-(2-fluorophenyl)-8-isobutyl-8-methyl-1,6-naphthyridin-7(8H)-one

Step 1.5-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-3-chloro-2-(2-fluorophenyl)-8-isobutyl-8-methy-1,6-naphthridine-7(8H)-one

To a 50-mL round-bottomed flask was added 2,4-dimethylpentanenitrile(Intermediate I-4x, 0.577 g, 5.19 mmol) in toluene (8.65 mL). Thereaction mixture was cooled to 0° C. with a wet ice bath. Then potassiumbis(trimethylsilyl)amide (1.0M in THF) (5.84 mL, 5.84 mmol) was added tothe reaction mixture. The reaction mixture was allowed to stir at 0° C.while under an inert (N₂) atmosphere for 30 min. Then a heterogeneousmixture of 2,5-dichloro-6-(2-fluorophenyl)nicotinamide (Intermediate99B, 0.37 g, 1.29 mmol) in toluene (8.65 mL) was added to the reactionmixture. The resulting reaction mixture was allowed to stir 30 min,while the temperature was maintained at 0° C., then the ice bath wasremoved and the reaction mixture was allowed to warm to ambienttemperature over 30 min. The reaction mixture was quenched with 1N HCl(2 mL), then the mixture was diluted with EtOAc. The layers wereseparated and the aqueous layer was extracted with EtOAc (3×). Thecombined organic extracts were dried over MgSO₄, filtered andconcentrated in vacuo. The crude material was absorbed onto a plug ofsilica gel and purified by chromatography through a Redi-Sep pre-packedsilica gel column (40 g), eluting with a gradient of 0-30% 3:1EtOAc/EtOH in heptane, to afford5-chloro-2-(2-cyano-4-methylpentan-2-yl)-6-(2-fluorophenyl)nicotinamide(0.325 g, 0.90 mmol, 69.6% yield) as tan solid. m/z (ESI, +ve ion):360.2 (M+H)⁺.

To5-chloro-2-(2-cyano-4-methylpentan-2-yl)-6-(2-fluorophenyl)nicotinamide(0.277 g, 0.77 mmol) in a 50-mL round-bottomed flask was addedhydrochloric acid, 36.5-38.0% (3.96 mL, 46.2 mmol) dropwise. Thereaction mixture was stirred at 70° C. for 24 h. The reaction mixturewas allowed to cool to rt. The mixture was treated with sat. aq. NaHCO₃.Then 5N NaOH was added to neutralize the reaction mixture to pH 6. Theprecipitate formed was collected by filtration, dried in a vacuum ovenat 50° C. to give3-chloro-2-(2-fluorophenyl)-8-isobutyl-8-methyl-1,6-naphthyridine-5,7(6H,8H)-dione(230 mg, 0.63 mmol, 83% yield) as a tan solid. m/z (ESI, +ve ion): 361.3(M+H)⁺. The material was used directly in the following step, withoutfurther purification.

Step 2.5-((S)-4-Acryloyl-2-methylpiperazin-1-yl)-3-chloro-2-(2-fluorophenyl)-8-isobutyl-8-methyl-1,6-naphthyridin-7(8H)-one

To a 25-mL round-bottomed flask was added3-chloro-2-(2-fluorophenyl)-8-isobutyl-8-methyl-1,6-naphthyridine-5,7(6H,8H)-dione(120 mg, 0.33 mmol) in toluene (1.66 mL). Then N,N-diisopropylethylamine(0.58 mL, 3.33 mmol) and phosphorous oxychloride (0.15 mL, 1.66 mmol)was added to the reaction mixture. The resulting reaction mixture washeated and stirred at 80° C. for 2.5 h, while under an inert (N₂)atmosphere. The flask was removed from the heat bath and the crudemixture was concentrated in vacuo. The crude residue was diluted withtoluene (1.5 mL) and set aside.

To a 50-mL round-bottomed flask was added(S)-1-(3-methylpiperazin-1-yl)prop-2-en-1-one 2,2,2-trifluoroacetate(Intermediate I-7x, 0.686 g, 0.99 mmol) in toluene (l mL). Then DIPEA(1.4 mL) was added to the reaction mixture. To this mixture was addedthe previous crude mixture dropwise. After the addition, the reactionmixture was quenched with sat. aq. NH₄Cl. The layers were separated andthe aqueous layer was extracted with EtOAc (3×). The combined organicextracts were dried over Na₂SO₄, filtered and concentrated in vacuo. Thecrude material was purified by chromatography through a Redi-Seppre-packed silica gel column (40 g), eluting with a gradient of 0-40%3:1 EtOAc/EtOH in heptane, to afford(R)-5-((S)-4-acryloyl-2-methylpiperazin-1-yl)-3-chloro-2-(2-fluorophenyl)-8-isobutyl-8-methyl-1,6-naphthyridin-7(8H)-oneand(S)-5-((S)-4-acryloyl-2-methylpiperazin-1-yl)-3-chloro-2-(2-fluorophenyl)-8-isobutyl-8-methyl-,6-naphthyridin-7(8H)-one (50 mg, 0.05 mmol, 15.1% yield) as off-whitesolid. m/z (ESI, +ve ion): 497.3 (M+H)⁺.

Section 2. Synthesis of Intermediates Intermediates 99 A and B2,5-Dichloro-6-(2-fluorophenyl)nicotinamide

Step 1: 2,5-Dichloro-6-(2-fluorophenyl)nicotinic Acid

A mixture of 2,5,6-trichloronicotinic acid (1.03 g, 4.54 mmol,Combi-Blocks, San Diego, Calif.), palladium tetrakis (0.131 g, 0.114mmol), (2-fluorophenyl)boronic acid (0.699 g, 5.0 mmol, TCI America,Portland, Oreg.), and sodium carbonate (2M in water, 6.82 mL, 13.6 mmol)in 1,4-dioxane (11 mL) was sparged with nitrogen and heated to 80° C.for 1 h followed by 90° C. for 5 h. The reaction mixture was dilutedwith EtOAc (150 mL), washed with 1 N aqueous citric acid (2×100 mL); theorganic layer was separated, dried over anhydrous Na₂SO₄, andconcentrated in vacuo to give 2,5-dichloro-6-(2-fluorophenyl)nicotinicacid (Intermediate 99A, 1.27 g, 4.43 mmol, 97% yield) as an amber oil.m/z (ESI, +ve ion): 285.8 (M+H)⁺.

Step 2: 2,5-Dichloro-6-(2-fluorophenyl)nicotinamide

A solution of 2,5-dichloro-6-(2-fluorophenyl)nicotinic acid(Intermediate 99A, 1.27 g, 4.43 mmol) in sulfurous dichloride (13 mL,177 mmol) was stirred at 70° C. for 30 min. The reaction mixture wasconcentrated in vacuo to give a dark brown oil. The oil was dissolved in1,4-dioxane (8.9 mL) and treated with ammonium hydroxide (30% aq., 3.5mL, 89 mmol) and the mixture was stirred at rt for 5 min. The reactionmixture was diluted with EtOAc (150 mL), added to a separatory funnel,and washed with saturated, aqueous sodium bicarbonate (3×100 mL) theorganic layer was separated, dried over anhydrous Na₂SO₄, andconcentrated in vacuo. The crude product was purified by silica gelchromatography (eluent: 0-80% EtOAc/heptane) to provide crude product asan off-white solid. The solid was stirred in EtOH (8 mL) at rt for 15min and filtered to give 2,5-dichloro-6-(2-fluorophenyl)nicotinamide(Intermediate 99B. 0.449 g, 1.58 mmol, 36% yield) as a white solid. m/z(ESI, +ve ion): 284.8 (M+H)⁺.

Intermediate I-1x

2-Cyclopentylpropanenitrile

To a 100-mL round-bottomed flask was added 1-cyclopentane-acetonitrile(1.20 mL, 11.0 mmol, Combi-Blocks, San Diego, Calif., USA) and2,2′-bipyridine (0.17 g, 1.09 mmol, Strem Chemicals, Newburyport, Mass.,USA) in tetrahydrofuran (18.3 mL). The reaction mixture was cooled to−78° C. with a dry ice/acetone bath. Then lithium diisopropylamide (1.0Min THF/hexanes) (13.7 mL, 13.7 mmol) was added dropwise to the reactionmixture over 5 min. The reaction mixture was kept cold at −78° C. andstirred for 45 min. Then iodomethane (1.36 mL, 22.0 mmol, Sigma-Aldrich,St. Louis, Mo., USA) was added dropwise to the reaction mixture. After 5min, the ice bath was removed and the reaction mixture was allowed towarm to ambient temperature. The reaction mixture was quenched with theaddition of 1N HCl (12 mL), and stirred an additional 10 min. Then EtOAcwas added to the mixture. The layers were separated and the aqueouslayer was extracted with EtOAc. The aqueous layer was diluted withEtOAc, then extracted with EtOAc (2×). The combined organic extractswere dried over MgSO₄ and filtered. The filtrate was set-up fordistillation with a short-path condenser. The solvents were removed (hotplate temp set to 105° C.; distillates evaporated at 80° C.). Thetan-colored liquid (Intermediate-1x; 1.20 g) was used without furtherpurification. ¹H NMR (400 MHz, CHLOROFORM-d) δ 2.51-264 (m, 1H) 195 (brd, J=7.05 Hz, 2H) 1.77-1.89 (m, 3H) 1.65-1.76 (m, 3H) 1.36-1.64 (m, 6H)1.31 (d, J=7.05 Hz, 3H).

Intermediate I-2x 2-Cyclopropylpropanenitrile

2-Cyclopropylpropanenitrile

To a 100-mL round-bottomed flask was added cyclopropylacetonitrile (2.0mL, 24.66 mmol, Alfa Aesar, Tewkbury, Mass., USA) and 2,2′-bipyridine(0.385 g, 2.46 mmol) in tetrahydrofuran (24.6 mL). The reaction mixturewas cooled to −78° C. with a dry ice/acetone bath. Then lithiumdiisopropylamide (1.0M in THF/hexanes) (30.8 mL, 30.8 mmol) was addeddropwise to the reaction mixture over 5 min. The reaction mixture waskept cold at −78° C. and the mixture was allowed to stir for 45 min.Then iodomethane (1.54 mL, 24.7 mmol) was added dropwise to the reactionmixture and the reaction mixture was allowed to slowly warm to ambienttemperature overnight. The reaction mixture was quenched with theaddition of 1N HCl (25 mL) and the mixture was stirred and additional 10min. Then EtOAc was added to the mixture. The layers were separated andthe aqueous layer was extracted with EtOAc. The aqueous layer wasdiluted with EtOAc and extracted with EtOAc (2×). The combined organicextracts were dried over MgSO₄ and filtered. The organics wereconcentrated *(cold evaporation process, by lifting the round-bottomedflask out of the water bath) to leave the desired material. The mixturewas filtered through a fine-fritted scintered glass funnel and the tancrude organic filtrate (Intermediate-2x, 2.40 g) was collected and usedwithout further purification. ¹H NMR (400 MHz, DMSO-d₆) δ 2.41-2.49 (m,1H) 1.29 (d, J=7.05 Hz, 3H) 0.99-1.03 (m, 1H) 0.52-0.58 (m, 2H)0.24-0.34 (m, 2H).

Intermediate I-3x 2-Cyclobutylpropanenitrile

2-Cyclobutylpropanenitrile

To a 100-mL round-bottomed flask was added 2-cyclobutylacetonitrile(2.00 mL, 21.0 mmol, Enamine Ltd., Monmouth, N.J., USA) and2,2′-bipyridine (0.328 g, 2.10 mmol) in tetrahydrofuran (21.0 mL). Thereaction mixture was cooled to −78° C. with a dry ice/acetone bath. Thenlithium diisopropylamide (1.0M in THF/hexanes) (26.3 mL, 26.3 mmol) wasadded dropwise to the reaction mixture over 5 min. The reaction mixturewas kept cold at −78° C. then the mixture was allowed to stir for 45min. Then iodomethane (1.31 mL, 21.0 mmol) was added dropwise to thereaction mixture and the reaction mixture was allowed to slowly warm toambient temperature overnight. The reaction mixture was quenched withthe addition of 1N HCl (22 mL) and stirred and additional 10 min. ThenEtOAc was added to the mixture. The layers were separated and theaqueous layer was extracted with EtOAc. The aqueous layer was dilutedwith EtOAc and extracted with EtOAc (2×). The combined organic extractswere dried over MgSO₄ and filtered. The organics were concentrated*(cold evaporation process, by lifting the round-bottomed flask out ofwater bath, during the evaporation) to leave the desired material. Themixture was filtered through a fine-fritted funnel and the tan crudeorganic filtrate (Intermediate-3x, 2.10 g) was collected and was usedwithout further purification. ¹H NMR (400 MHz, DMSO-d₆) δ 2.81 (quin,J=7.26 Hz, 1H) 2.26-2.45 (m, 1H) 1.96-2.02 (m, 2H) 1.74-1.83 (m, 4H)1.09 (d, J=7.05 Hz, 3H).

Intermediate I-4x 2,4-Dimethylpentanenitrile

2,4-Dimethylpentanenitrile

A mixture of isocapronitrile (2.8 g, 28.8 mmol, TCI America, Portland,Oreg., USA) and 2,2′-bipyridine (0.450 g, 2.88 mmol) in tetrahydrofuran(14.4 mL) was added to a cold (−78° C.) stirred solution of lithiumdiisopropylamide (2.0M in THF/hexanes) (18.0 mL, 36.0 mmol). After theaddition was complete, the reaction mixture was stirred at −78° C. for10 min, then iodomethane (3.59 mL, 57.6 mmol) was added dropwise. Thereaction mixture was stirred at −78° C. for 1 h. The reaction wasquenched with the addition of 1.0 N HCl (50 mL), then the mixture wasallowed to warm to room temperature. Then THF was evaporated and themixture was extracted with tBuOMe (3×50 mL). The combined organicextracts were washed with 1.0 N HCl (20 mL) and sat. NaHCO₃(30 mL). Theorganic layer was dried over Na₂SO₄ and evaporated. The residue wasdistilled (150° C.) at atmospheric pressure. The remaining material darkliquid (Intermediate-4x, 0.582 g), was collected and was used withoutfurther purification.

Intermediate I-5x 2,2-Dicyclopropylacetonitrile

2,2-Dicyclopropylacetamide

A mixture of 2,2-dicyclopropylacetic acid (2.0 g, 14.2 mmol) and thionylchloride (2.60 mL, 35.7 mmol) was heated to reflux for 30 min. Theexcess of thionyl chloride was removed in vacuo, and the acyl chloridewas used in the next step as is.

A solution of ammonium hydroxide (10 mL, 10.0 mmol) was cooled to 0° C.with a wet ice/water bath and a solution of the acyl chloride in THF (6mL) was carefully added dropwise. The reaction mixture was stirred at rtin n open flask overnight to remove excess ammonia. The whiteprecipitate was filtered, washed with cold water, and azeotropicallydried with toluene to afford 2,2-dicyclopropylacetamide (1.55 g, 11.4mmol, 78% yield) as an off-white solid that was used without furtherpurification. ¹H NMR (400 MHz, DMSO-d₆) δ 6.89 (br s, 1H), 6.49 (br s,1H), 0.79 (td, J=8.7, 4.0 Hz, 2H), 0.65 (d, J=9.1 Hz, 1H), 0.24-0.34 (m,2H), 0.15 (dt, J=8.4, 4.4 Hz, 2H), −0.09-0.05 (m, 4H). m/z (ESI, +veion): 140 (M+H)⁺.

Step 2. 2,2-Dicyclopropylacetonitrile

To a solution of 2,2-dicyclopropylacetamide (1.55 g, 11.1 mmol) in THF(25 mL) while at 0° C., was added pyridine (1.80 mL, 22.2 mmol). Theresulting mixture was stirred at 0° C. for 1 h. Then TFAA (7.86 mL, 55.7mmol) was added in one portion, then the mixture was allowed to stir anadditional 15 min, while at 0° C. The mixture was carefully basifiedwith sat. NaHCO₃ and extracted with DCM. The combined organics weredried over Na₂SO₄, filtered, and the solvents were evaporated to afford2,2-dicyclopropylacetonitrile (0.90 g, 7.43 mmol, 66.7% yield) as ayellow oil. ¹H NMR (400 MHz, DMSO-d₆) δ 2.20 (t, J=8.0 Hz, 1H),0.99-1.19 (m, 2H), 0.48-0.65 (m, 4H), 0.24-0.42 (m, 4H).

Intermediate I-6x 2,3-Dihydro-1H-indene-2-carbonitrile

Step 1. 2,3-Dihydro-1H-indene-2-carboxamide

A mixture of 2-indancarboxylic acid (1.0 g, 6.17 mmol, Oakwood Products,Estill, S.C., USA) and thionyl chloride (1.12 mL, 15.4 mmol) was heatedto reflux for 30 min. The excess thionyl chloride was removed in vacuo,and the acyl chloride was used in the next step as is.

A solution of ammonium hydroxide (5.0 mL, 5.00 mmol) was cooled to 0°C., then a solution of the acyl chloride in THF (6 mL) was carefullyadded dropwise. The reaction mixture was stirred overnight, while at rtin an open flask to remove excess ammonia. The white precipitate wasfiltered, washed with cold water, and azeotropically dried with tolueneto afford 2,3-dihydro-1H-indene-2-carboxamide (0.835 g, 5.18 mmol, 84%yield) as a white solid which was used as is. ¹H NMR (400 MHz, DMSO-d₆)δ 7.40 (br s, 1H), 7.17-7.22 (m, 2H), 7.12 (dd, J=5.5, 32 Hz, 2H), 6.85(br s, 1H). 3.10-3.19 (m, 1H), 3.02-3.08 (m, 4H). m/z (ESI, +ve ion):162.0 (M+H)⁺.

Step 2. 2,3-Dihydro-1H-indene-2-carbonitrile

To a solution of 2,3-dihydro-1H-indene-2-carboxamide (0.835 g, 5.18mmol) in THF (25 mL) at 0° C. was added pyridine (0.83 mL, 10.3 mmol).The resulting mixture was stirred at 0° C. for 1 h. Then TFAA (3.66 mL,25.9 mmol) was added in one portion then the mixture was allowed to stiran additional 15 min, while at 0° C. The mixture was carefully basifiedwith sat. NaHCO₃ and the aqueous layer was extracted with DCM. Thecombined organic extracts were washed with sat. NH₄Cl, dried overNa₂SO₄, filtered and concentrated in vacuo. The crude material waspurified by silica-gel chromatography, with a gradient of 0-30% EtOAc inheptane to afford 2,3-dihydro-1H-indene-2-carbonitrile (0.701 g, 4.90mmol, 95% yield) was a colorless oil, that was azeotropically dried withtoluene. ¹H NMR (400 MHz, DMSO-d₆) δ 7.24-7.32 (m, 2H), 7.20 (dd, J=5.6,3.3 Hz, 2H), 3.50-3.63 (m, 1H), 3.33-3.37 (m, 1H), 3.29 (s, 1H), 3.16(d, J=6.8 Hz, 1H), 3.13 (d, J=7.0 Hz, 1H). m/z (ESI, +ve ion): 144.0(M+H)⁺.

Intermediate I-7x (S)-1-(3-Methylpiperazin-1-yl)prop-2-en-1-one2,2,2-trifluoroacetate

Step 1 (S)-tert-Butyl 4-acryloyl-2-methylpiperazine-1-carboxylate

Acryloyl chloride (1.34 mL, 16.5 mmol) was added to a solution of(S)-1-Boc-2-methyl-piperazine (3.00 g, 15.0 mmol, Boc Sciences, Shirley,N.Y.) in THF (30 mL) at −10° C., and the resulting mixture was stirredat −10° C. for 5 min. Triethylamine (6.26 mL, 44.9 mmol) was then slowlyadded, and the resulting mixture was stirred at −10° C. for 15 min, thenallowed to warm to rt. The reaction mixture was partitioned betweenEtOAc and saturated aqueous NaHCO3. The aqueous layer was extracted withEtOAc (3×), and the organic layers were then combined, dried over MgSO4,filtered, and concentrated in vacuo. Chromatographic purification of theresidue (silica gel, 0-100% EtOAc in heptane) furnished (S)-tert-butyl4-acryloyl-2-methylpiperazine-1-carboxylate: 1H NMR (400 MHz, DMSO-d6) δ6.72-6.85 (m, 1H) 6.10-6.18 (m, 1H) 5.68-5.76 (m, 1H) 4.08-4.32 (m, 2H)3.68-4.03 (m, 2H) 2.86-3.14 (m, 2H) 2.66-2.80 (m, 1H) 1.38-1.43 (s, 9H)0.96-1.04 (m, 3H). m/z (ESI, +ve) 277.3 (M+Na)+.

Step 2. (S)-1-(3-Methylpiperazin-1-yl)prop-2-en-1-one2,2,2-trifluoroacetate

A mixture of (S)-tert-butyl 4-acryloyl-2-methylpiperazine-1-carboxylate(3.21 g, 12.6 mmol) and TFA (4.7 mL, 63.1 mmol) in DCM (16 mL) wasstirred at rt for 24 h. The reaction mixture was then concentrated invacuo to give (S)-1-(3-methylpiperazin-1-yl)prop-2-en-1-one2,2,2-trifluoroacetate: 1H NMR (400 MHz, DMSO-d6) δ 8.70-8.99 (m, 1H)6.74-6.91 (min, 1H) 6.12-6.26 (m, 1H) 5.70-5.84 (m, 1H) 4.25-4.44 (m,1H) 4.07-4.25 (m, 1H) 3.49-3.53 (m, 1H) 3.22-3.32 (m, 2H) 2.92-3.08 (m,2H) 1.14-1.29 (m, 3H). m/z (ESI, +ve) 155.1 (M+H)+.

Intermediate 160 1-((2R,5S)-2,5-Dimethylpiperazin-1-yl)prop-2-en-1-one2,2,2-trifluoroacetate

To a solution of (2S,5R)-1-Boc-2,5-dimethylpiperazine (3.5 g, 16.3 mmol,Astatech) in dichloromethane (35 ml), anhydrous triethylamine (4.59 ml,32.7 mmol) was added and the mixture was cooled to 0° C. in ice-waterbath. Acryloyl chloride (1.46 ml, 18.0 mmol) was added dropwise over −5min at which point the reaction mixture became yellow and viscous andformation of a white precipitate was observed. Water (10 ml) was addedand the mixture was removed from the ice bath and was allowed to stirfor 10 min. The organic layer was separated, quickly washed with 2N HCl(40 ml), water and brine, filtered through pad of MgSO₄ and concentratedto afford crude tert-butyl(2S,5R)-4-acryloyl-2,5-dimethylpiperazine-1-carboxylate (4.4 g) asyellow oil. This material was redissolved in DCM (40 ml) and TFA (12.6ml, 163 mmol) was added and the mixture was stirred at rt for 4 h atwhich point complete deprotection was observed. The mixture wasconcentrated under reduced pressure and dried under vacuum to afford1-((2R,5S)-2,5-dimethylpiperazin-1-yl)prop-2-en-1-one2,2,2-trifluoroacetate (Intermediate 160, ˜9 g) as an oil. Analysis byqNMR using benzyl benzoate as internal standard showed 40.2 wt % purity.The material was used without further purification. ¹H NMR (400 MHz,DMSO-d₆) δ 9.03 (br s, 1H), 8.95 (br s, 1H), 6.77 (dd, J=0.6, 16.8 Hz,1H), 6.16 (dd, J=2.3, 16.8 Hz, 1H), 5.74 (dd, J=2.3, 10.6 Hz, 1H),4.69-4.56 (m, 1H), 4.09-3.94 (m, 1H), 3.70-3.57 (m, 1H), 3.45-3.19 (m,2H), 2.98-3.09 (m, 1H), 1.25 (d, J=6.8 Hz, 3H), 1.20 (d, J=6.8 Hz, 3H).m/z (ESI, +ve ion): 169.3 (M+H).

TABLE 3 Separated Compounds Table Racemic SM/Separation Ex. # ChemicalStructure Name Conditions 1-1-1

5-((2S,5R)-4-acryloyl-2,5- dimethylpiperazin-1-yl)-3-chloro-8-cyclopentyl-2-(2- fluorophenyl)-8-methyl-1,6-naphthyridin-7(8H)- one 1-1/SFC (Column: OX (250 × 21 mm, 5 μm) F =70 mL/min, 35% methanol, 65% carbon dioxide, 102 bar) 1-1-2

5-((2S,5R)-4-acryloyl-2,5- dimethypiperazin-1-yl)-3-chloro-8-cyclopentyl-2-(2- fluorophenyl)-8-methyl-1,6-naphthyridin-7(8H)- one 1-1/SFC (Column: OX (250 × 21 mm, 5 μm) F =70 mL/min, 35% methanol, 65% carbon dioxide, 102 bar) 1-2-1

5-((2S,5R)-4-acroyl-2,5- dimethylpiperazin-1-yl)-3-chloro-8-cyclopropyl-2- (2-fluorophenyl)-8- methyl-1,6-naphthyridin-7(8H)-one 1-1/SFC (Column: OD (250 × 21 mm, 5 μm) F = 80 mL/min, 25%methanol, 75% carbon dioxide, 102 bar) 1-2-2

5-((2S,5R)-4-acryloyl-2,5- dimethyliperazin-1-yl)-3-chloro-8-cyclopropyl-2- (2-fluorophenyl)-8- methyl-1,6-naphthyridin-7(8H)-one 1-1/SFC (Column: OD (250 × 21 mm, 5 μm) F = 80 mL/min, 25%methanol, 75% carbon dioxide, 102 bar) 1-3-1

5-((2S,5R)-4-acryloyl-2,5- dimethylpiperazin-1-yl)-3-chloro-8-cyclobutyl-2-(2- fluorophenyl)-8-methyl-1,6-naphthyridin-7(8H)- one 1-1/SFC (Column: OX (250 × 21 mm, 5 μm) F =75 mL/min, 40% methanol, 60% carbon dioxide, 102 bar) 1-3-2

5-((2S,5R)-4-acryloyl-2,5- dimethylpiperazin-1-yl)-3-chloro-8-cyclobutyl-2-(2- fluorophenyl)-8-methyl-1,6-naphthyridin-7(8H)- one 1-1/SFC (Column: OX (250 × 21 mm, 5 μm) F =75 mL/min, 40% methanol, 60% carbon dioxide, 102 bar) 1-4-1

5-((2S,5R)-4-acryloyl-2,5- dimethylpiperazin-1-yl)-3-chloro-8-ethyl-2-(2- fluorophenyl)-8-methyl- 1,6-naphthyridin-7(8H)- one1-1/SFC (Column: OD (250 × 21 mm, 5 μm) F = 90 mL/min, 10% methanol, 90%carbon dioxide, 102 bar) 1-4-2

5-((2S,5R)-4-acryloyl-2,5- dimethylpiperazin-1-yl)-3-chloro-8-ethyl-2-(2- fluorophenyl)-8-methyl- 1,6-naphthyridin-7(8H)- one1-1/SFC (Column: OD (250 × 21 mm, 5 μm) F = 90 mL/min, 10% methanol, 90%carbon dioxide, 102 bar) 1-5-1

5-((2S,5R)-4-acryloyl-2,5- dimethylpiperazin-1-yl)-3-chloro-2-(2-fluorophenyl)- 8-methyl-8-phenyl-1,6- naphthyridin-7(8H)-one1-1/SFC (Column: OX (150 × 30 mm, 5 μm) F = 180 mL/min, 25% methanol,75% carbon dioxide, 102 bar) 1-5-2

5-((2S,5R)-4-acryloyl-2,5- dimethylpiperazin-1-yl)-3-chloro-2-(2-fluorophenyl)- 8-methyl-8-phenyl-1,6- naphthyridin-7(8H)-one1-1/SFC (Column: OX (150 × 30 mm, 5 μm) F = 180 mL/min, 25% methanol,75% carbon dioxide, 102 bar) 1-7-1

5′-((2S,5R)-4-acryloyl- 2,5-dimethylpiperazin-1- yl)-3′-chlroo-2′-(2-fluorophenyl)-2,3- dihydro-7′H-spiro[indene- 1,8′-[1,6]naphthyridin]-7′-one 1-1/SFC (Column: OX (250 × 21 mm, 5 μm) F = 80 mL/min, 40% methanol,60% carbon dioxide, 90 bar) 1-7-2

5′-((2S,5R)-4-acryloyl- 2,5-dimethylpiperazin-1- yl)-3′-chloro-2′-(2-fluorophenyl)-2,3- dihdyro-7′H-spiro[indene- 1,8′-[1,6]naphthyridin]-7′-one 1-1/SFC (Column: OX (250 × 21 mm, 5 μm) F = 80 mL/min, 40% methanol,60% carbon dioxide, 90 bar) 1-8-1

5-((2S,5R)-4-acrloyl-2,5- dimethylpiperazin-1-yl)-3-chloro-2-(2-fluorophenyl)- 8-isobutyl-8-methyl-1,6-naphthyridin-7(8H)-one 1-1/SFC (Column: OX (250 × 21 mm, 5 μm) F = 90mL/min, 20% methanol, 80% carbon dioxide, 102 bar) 1-8-2

5-((2S,5R)-4-acryloyl-2,5- dimethylpiperazin-1-yl)-3-chloro-2-(2-fluorophenyl)- 8-isobutyl-8-methyl-1,6-naphthyridin-7(8H)-one 1-1/SFC (Column: OX (250 × 21 mm, 5 μm) F = 90mL/min, 20% methanol, 80% carbon dioxide, 102 bar) 3-1-1

5-((S)-4-acryloyl-2- methylpiperazin-1-yl)-3- chloro-2-(2-fluorophenyl)-8-isobutyl-8-methyl-1,6- naphthyridin-7(8H)-one 1-1/SFC (Column: OD-H(21 × 250 mm, 5 μm) F = 80 mL/min, 25% methanol, 75% carbon dioxide, 101bar) 3-1-2

5-((S)-4-acryloyl-2- methylpiperazin-1-yl)-3- chloro-2-(2-fluorophenyl)-8-isobutyl-8-methyl-1,6- naphthyridin-7(8H)-one 1-1/SFC (Column: OD-H(21 × 250 mm, 5 μm) F = 80 mL/min, 25% methanol, 75% carbon dioxide, 101bar)

Section 3. Supporting Data

TABLE 4 Mass spectroscopy and NMR analytical Data LRMS: (ESI Ex. # + veion) m/z NMR 1-1 523.1 ¹H NMR (400 MHz, DMSO-d₆) δ 8.21 (br s, 1H)7.46-7.57 (m, 2H) 7.28-7.36 (m, 2H) 6.65-6.83 (m, 1H) 6.11 (br d, J =17.21 Hz, 1H) 5.68 (br d, J = 9.95 Hz, 1 H) 4.73-5.11 (m, 1H) 4.64 (brs, 1H) 4.31-4.51 (m, 1H) 3.75-3.96 (m, 1H) 3.43 (br d, J = 12.23 Hz, 1H)2.14 (br s, 1 H) 1.42 (br s, 3H) 1.24-1.38 (m, 10 H) 1.18 (br d, J =6.84 Hz, 3H) 1.03-1.13 (m, 1H) 0.98 (br s, 1H). 1-1-1 523.2 ¹H NMR (400MHz, DMSO-d₆) δ 8.16- 8.27 (m, 1H) 7.46-7.56 (m, 2H) 7.28- 7.35 (m, 2H)6.69-6.80 (m, 1H) 6.08- 6.15 (m, 1H) 5.65-5.71 (m, 1H) 5.00 (br s, 1H)4.66 (br s, 1H) 4.36 (br s, 1H) 3.95- 4.21 (m, 1H) 3.76-3.95 (m, 2H)3.68 (br s, 1H) 2.15 (br s, 1H) 1.27-1.43 (m, 10 H) 1.14-1.24 (m, 3H)1.06 (br s, 1H) 0.98 (br s, 2H) 1-1-2 523.2 ¹H NMR (400 MHz, DMSO-d₆) δ7.99 (d, J = 10.78 Hz, 1H) 7.53-7.65 (m, 2H) 7.35- 7.45 (m, 2H)6.72-6.91 (m, 1H) 6.12- 6.22 (m, 1H) 5.71-5.79 (m, 1H) 4.87 (br s, 1H)4.61-4.75 (m, 1H) 4.55 (br s, 1H) 4.19-4.31 (m, 1H) 3.79-3.94 (m, 1H)3.38-3.59 (m, 1H) 2.97-3.14 (m, 1H) 2.22 (br s, 1H) 1.47-1.59 (m, 2H)1.37- 1.45 (m, 7H) 1.35 (s, 4H) 1.26 (br s, 3H) 1-2 495.0 ¹H NMR (400MHz, DMSO-d₆) δ 7.71- 7.78 (m, 1H) 7.30-7.41 (m, 2H) 7.12- 7.20 (m, 2H)6.48-6.68 (m, 1H) 5.89- 6.01 (m, 1H) 5.47-5.57 (m, 1H) 4.41- 4.69 (m,1H) 3.98-4.37 (m, 1H) 3.60- 3.86 (m, 1H) 3.32-3.52 (m, 1H) 2.87- 2.95(m, 1H) 1.19 (br d, J = 3.52 Hz, 3H) 1.02-1.07 (m, 6H) 0.98-1.01 (m, 2H)0.12-0.24 (m, 2H) −0.13-0.04 (m, 1H) −0.31-−0.15 (m, 1H) 1-2-1 495.1 ¹HNMR (400 MHz, DMSO-d₆) δ 7.83 (d, J = 9.95 Hz, 1H) 7.40-7.50 (m, 2H)7.23- 7.29 (m, 2H) 6.59-6.77 (m, 1H) 6.00- 6.07 (m, 1H) 5.59-5.65 (m,1H) 4.73 (br s, 1H) 4.60 (br s, 1H) 4.55 (br s, 1H) 4.42 (br s, 1H)4.06-4.20 (m, 1H) 3.36 (br d, J = 14.31 Hz, 1H) 1.28 (s, 6H) 1.04-1.22(m, 4H) 0.23-0.34 (m, 2H) 0.00 (br s, 1 H) −0.15 (br s, 1H) 1-2-2 495.1¹H NMR (400 MHz, DMSO-d₆) δ 8.07 (br s, 1H) 7.31-7.41 (m, 2H) 7.13-7.20(m, 2H) 6.54-6.66 (m, 1H) 5.96 (br d, J = 16.59 Hz, 1H) 5.52 (br d, J =10.37 Hz, 1 H) 4.91 (br s, 1H) 4.53 (br s, 1H) 3.95 (br d, J = 13.68 Hz,1H) 3.75 (br s, 1H) 3.68 (br d, J = 13.06 Hz, 1H) 1.19 (s, 3H) 0.91-1.08 (m, 6H) 0.87 (br s, 2H) 0.12-0.22 (m, 2H) −0.05-0.05 (m, 1H)−0.26-−0.15 (m, 1H) 1-3 509.1 ¹H NMR (400 MHz, DMSO-d₆) δ 7.96- 8.06 (m,1H) 7.53-7.64 (m, 2H) 7.33- 7.44 (m, 2H) 6.72-6.90 (m, 1H) 6.18 (dt, J =16.74, 2.00 Hz, 1H) 5.68-5.78 (m, 1 H) 4.96-5.23 (m, 1H) 4.59-4.92 (m, 1H) 4.34-4.57 (m, 1H) 4.03-4.32 (m, 1 H) 3.71-3.95 (m, 1H) 3.40-3.67 (m,1 H) 2.61-2.72 (m, 1H) 1.58-1.84 (m, 5 H) 1.48 (br d, J = 8.29 Hz, 1H)1.35 (s, 5 H) 1.13-1.30 (m, 3H) 0.97-1.12 (m, 1 H) 1-3-1 509.1 ¹H NMR(400 MHz, DMSO-d₆) δ 8.30 (br s, 1H) 7.53-7.64 (m, 2H) 7.34-7.43 (m, 2H)6.76-6.88 (m, 1H) 6.18 (br d, J = 16.59 Hz, 1H) 5.75 (br d, J = 9.95 Hz,1 H) 5.09 (br s, 1H) 4.76 (br s, 1H) 4.45 (br s, 1H) 4.17 (br d, J =12.85 Hz, 1H) 3.98 (br s, 1H) 3.90 (br d, J = 13.27 Hz, 1H) 2.61-2.70(m, 1H) 1.57-1.84 (m, 5H) 1.48 (br d, J = 8.71 Hz, 1H) 1.35 (s, 3H)1.14-1.29 (m, 4H) 0.98-1.12 (m, 2H) 1-3-2 509.1 ¹H NMR (400 MHz,DMSO-d₆) δ 8.02 (d, J = 12.23 Hz, 1H) 7.56-7.65 (m, 2H) 7.37- 7.44 (m,2H) 6.74-6.91 (m, 1H) 6.18 (br d, J = 16.59 Hz, 1H) 5.73-5.79 (m, 1 H)4.85 (br s, 1H) 4.67 (br s, 1H) 4.53 (br s, 1H) 4.27 (br d, J = 13.48Hz, 1H) 3.35- 3.59 (m, 2H) 2.64-2.74 (m, 1H) 1.81 (br d, J = 8.50 Hz,1H) 1.75 (br s, 2H) 1.59- 1.72 (m, 2H) 1.49 (br d, J = 7.05 Hz, 2H)1.30-1.43 (m, 8H) 1-4 483.1 ¹H NMR (400 MHz, DMSO-d₆) δ 8.01- 8.31 (m,1H) 7.52-7.64 (m, 2H) 7.34- 7.44 (m, 2H) 6.73-6.88 (m, 1H) 6.18 (br d, J= 16.59 Hz, 1H) 5.70-5.78 (m, 1H) 4.78 (br d, J = 17.00 Hz, 1H)4.09-4.25 (m, 1H) 3.81-3.94 (m, 1H) 3.62 (br dd, J = 13.16, 3.42 Hz, 1H)3.29-3.33 (m, 1 H) 1.79-1.96 (m, 2H) 1.40 (br d, J = 4.35 Hz, 4H)1.24-1.31 (m, 2H) 1.14-1.24 (m, 3H) 0.98-1.12 (m, 1H) 0.65-0.85 (m, 3H)1-4-1 483.2 ¹H NMR (400 MHz, DMSO-d₆) δ 8.10 (br d, J = 10.57 Hz, 1H)7.53-7.64 (m, 2H) 7.35-7.43 (m, 2H) 6.71-6.89 (m, 1H) 6.12-6.21 (m, 1H)5.70-5.78 (m, 1H) 4.80 (br s, 1H) 4.51 (br s, 1H) 4.20 (br d, J = 13.68Hz, 1H) 3.86 (br d, J = 14.10 Hz, 1 H) 3.62 (br d, J = 13.27 Hz, 1H)1.78-2.00 (m, 2H) 1.39 (s, 6H) 1.08-1.31 (m, 4H) 0.63-0.81 (m, 3H) 1-4-2483.2 ¹H NMR (400 MHz, DMSO-d₆) δ 8.26 (s, 1H) 7.54-7.63 (m, 2H)7.35-7.42 (m, 2 H) 6.75-6.87 (m, 1H) 6.18 (br d, J = 16.79 Hz, 1H)5.71-5.77 (m, 1H) 4.97 (br s, 1 H) 4.74 (br s, 1H) 4.45 (br s, 1H) 4.05-4.24 (m, 1H) 3.82-3.97 (m, 1H) 3.74 (br d, J = 12.65 Hz, 1H) 1.84 (q, J= 7.05 Hz, 2 H) 1.40 (s, 3H) 1.21-1.32 (m, 3H) 1.12- 1.18 (m, 1H) 1.08(br d, J = 5.80 Hz, 2H) 0.67-0.84 (m, 3H) 1-5 531.0 ¹H NMR (400 MHz,DMSO-d₆) δ 7.44- 7.58 (m, 2H) 7.22-7.33 (m, 2H) 7.02- 7.21 (m, 4H) 6.92(br d, J = 7.67 Hz, 1H) 6.89 (br d, J = 7.67 Hz, 1H) 6.47-6.64 (m, 1H)5.97 (br d, J = 15.76 Hz, 1H) 5.44- 5.59 (m, 1H) 4.34-4.60 (m, 1H) 3.97-4.21 (m, 1H) 3.39-3.64 (m, 1H) 2.84- 3.09 (m, 1H) 1.50-1.58 (m, 3H)1.23- 1.38 (m, 1H) 1.09 (s, 4H) 0.92-0.99 (m, 1H) 0.87 (br d, J = 6.01Hz, 1H) 0.65- 0.76 (m, 1H) 1-5-1 531.1 ¹H NMR (400 MHz, DMSO-d₆) δ 8.14(br s, 1H) 7.44-7.55 (m, 2H) 7.27 (t, J = 8.39 Hz, 2H) 7.01-7.15 (m, 3H)6.92 (br d, J = 7.46 Hz, 2H) 6.56 (br dd, J = 16.59, 10.57 Hz, 1H) 5.96(br dd, J = 16.69, 3.01 Hz, 1H) 5.54 (br d, J = 10.57 Hz, 1H) 4.91 (brs, 1H) 4.33 (br s, 1H) 3.86 (br s, 2H) 3.36-3.66 (m, 2H) 2.62-2.79 (m,1H) 1.54 (s, 3H) 1.09 (br s, 3H) 0.00 (br s, 2 H) 1-5-2 531.1 ¹H NMR(400 MHz, DMSO-d₆) δ 7.74- 7.90 (m, 1H) 7.35-7.50 (m, 2H) 7.19- 7.29 (m,2H) 6.99-7.12 (m, 3H) 6.86 (br d, J = 7.26 Hz, 2H) 6.43-6.63 (m, 1H)5.92 (br t, J = 16.17 Hz, 1H) 5.52 (br dd, J = 9.43, 7.15 Hz, 1H)4.20-4.56 (m, 2H) 4.03-4.18 (m, 1H) 3.78-3.98 (m, 1H) 3.34-3.61 (m, 1H)2.87-3.08 (m, 1H) 1.80-2.09 (m, 1H) 1.54 (s, 3H) 1.25 (br d, J = 13.48Hz, 2H) 1.00-1.18 (m, 1H) 0.05 (br s, 2H) 1-6 509.1 ¹H NMR (400 MHz,DMSO-d₆) δ 8.13 (d, J = 5.60 Hz, 1H) 7.55-7.64 (m, 2H) 7.36- 7.43 (m,2H) 6.70-6.88 (m, 1H) 6.11- 6.21 (m, 1H) 5.70-5.78 (m, 1H) 4.77 (br d, J= 15.96 Hz, 1H) 4.49 (br s, 1H) 4.11- 4.24 (m, 1H) 3.75-3.93 (m, 1H)3.60- 3.75 (m, 1H) 3.52-3.59 (m, 1H) 1.93 (br s, 3H) 1.65 (br s, 3H)1.42-1.60 (m, 2H) 1.31 (br s, 4H) 1.09-1.19 (m, 4H) 1-7 543.1 ¹H NMR(400 MHz, CDCl₃) δ 7.77-7.93 (m, 1H), 7.41-7.56 (m, 2H), 6.91-7.25 (m,5H), 6.50-6.87 (m, 2H), 6.35-6.46 (m, 1H), 5.77-5.85 (m, 1H), 4.26-5.26(m, 3H), 2.89-4.06 (m, 7H), 1.33-1.66 (m, 6H) 1-7-1 543.0 ¹H NMR (400MHz, DMSO-d₆) δ 8.31 (s, 1H), 7.43-7.54 (m, 2H), 7.19-7.32 (m, 3H),7.08-7.19 (m, 1H), 6.99 (br t, J = 7.36 Hz, 1H), 6.89 (br d, J = 7.67Hz, 1H), 6.72-6.85 (m, 1H), 6.14 (br d, J = 16.59 Hz, 1H), 5.67- 5.73(m, 1H) 3.29-5.39 (m, 6H), 3.02-3.15 (m, 2H), 2.86-3.01 (m, 1H), 2.73(br dd, J = 5.80, 12.85 Hz, 1H), 1.04-1.29 (m, 6H) 1-7-2 543.1 ¹H NMR(400 MHz, DMSO-d₆) δ 8.17 (d, J = 10.99 Hz, 1H), 7.49-7.62 (m, 2H),7.26- 7.40 (m, 3H), 7.10-7.22 (m, 2H), 6.77-6.97 (m, 2H), 6.20 (td, J =2.62, 16.74 Hz, 1H), 5.77 (br d, J = 10.37 Hz, 1H), 3.48-5.05 (m, 6H),2.98-3.23 (m, 3H), 2.65-2.81 (m, 1H), 1.13-1.61 (m, 6H) 1-8 511.3 ¹H NMR(400 MHz, DMSO-d₆) δ 8.08- 8.29 (m, 1H), 7.53-7.66 (m, 2H), 7.33-7.46(m, 2H), 6.71-6.92 (m, 1H), 6.18 (br d, J = 16.59 Hz, 1H), 5.71-5.79 (m,1H), 3.53- 5.06 (m, 6H), 1.59-1.86 (m, 3H), 1.44-1.45 (m, 3H), 0.98-1.34(m, 6H), 0.64-0.80 (m, 6H) 1-8-1 511.3 ¹H NMR (400 MHz, CHLOROFORM-d) δ7.77 (d, J = 8.50 Hz, 1H), 7.42-7.59 (m, 2H), 7.29 (t, J = 7.57 Hz, 1H),7.18 (t, J = 9.12 Hz, 1H), 6.44-6.69 (m, 1H), 6.30- 6.43 (m, 1H),5.72-5.83 (m, 1H), 4.07-5.17 (m, 3H), 3.03-3.67 (m, 3H), 1.67-1.96 (m,3H), 1.58 (s, 3H), 1.41 (br dd, J = 6.53, 13.16 Hz, 3H), 1.21-1.37 (m,3H), 0.66- 0.79 (m, 6H) 1-8-2 511.3 ¹H NMR (400 MHz, CHLOROFORM-d) δ7.68-7.76 (m, 1H), 7.42-7.57 (m, 2H), 7.26-7.35 (m, 1H), 7.18 (t, J =9.23 Hz, 1H), 6.45-6.67 (m, 1H), 6.29-6.43 (m, 1H), 5.71-5.86 (m, 1H),3.29-5.46 (m, 6H), 1.82-1.92 (m, 1H), 1.72-1.81 (m, 2H), 1.61 (s, 3H),1.36 (br d, J = 6.63 Hz, 3H), 1.18- 1.31 (m, 3H), 0.74-0.82 (m, 6H) 2-1521.6 ¹H NMR (400 MHz, DMSO-d₆) δ 8.08- 8.20 (m, 1H), 7.54-7.67 (m, 2H),7.32- 7.47 (m, 2H), 6.74-6.93 (m, 1H), 6.12- 6.24 (m, 1H), 5.75 (br d, J= 10.0 Hz, 1H), 4.74-5.03 (m, 1H), 4.39-4.59 (m, 1H), 4.12-4.30 (m, 1H),3.82-3.95 (m, 1H), 3.56-3.76 (m, 1H), 1.29-1.41 (m, 5H), 1.22-1.28 (m,2H), 1.15-1.20 (m, 2H), 0.32-0.60 (m, 8H) 2-2 542.6 ¹H NMR (400 MHz,DMSO-d₆) δ 8.23 (s, 1 H), 7.55 (dt, J = 13.0, 6.7 Hz, 2H), 7.29- 7.40(m, 2H), 7.07-7.24 (m, 4H), 6.81 (br dd, J = 16.2, 10.2 Hz, 1H), 6.20(br dd, J = 19.5, 2.9 Hz, 1H), 5.73-5.81 (m, 1H), 4.74-4.92 (m, 1H),4.53 (br d, J = 3.3 Hz, 1H), 4.21 (br d, J = 13.5 Hz, 1H), 3.91 (br d, J= 14.5 Hz, 1H), 3.72-3.84 (m, 2H), 3.53-3.70 (m, 3H), 3.36-3.45 (m, 1H),1.27 (br d, J = 6.2 Hz, 3H), 1.20 (br d, J = 6.6 Hz, 3H) 3-1 497.3 ¹HNMR (400 MHz, CHLOROFORM-d) δ 7.75 (br s, 1H), 7.43-7.57 (m, 2H), 7.26-7.34 (m, 1H), 7.18 (t, J = 9.12 Hz, 1H), 6.49-6.69 (m, 1H), 6.32-6.44(m, 1H), 5.78 (dd, J = 1.66, 10.57 Hz, 1H), 2.61-5.44 (m, 7H), 1.86-1.98(m, 1H), 1.62-1.85 (m, 2H), 1.59 (s, 3H), 1.36 (d, J = 6.84 Hz, 3H),0.77 (br d, J = 6.43 Hz, 6H) 3-1-1 497.3 ¹H NMR (400 MHz, CHLOROFORM-d)δ 7.73 (s, 1H), 7.42-7.58 (m, 2H), 7.29 (t, J = 7.57 Hz, 1H), 7.18 (t, J= 9.02 Hz, 1H), 6.46-6.70 (m, 1H), 6.35-6.43 (m, 1H), 5.79 (br d, J = 10.99 Hz, 1H), 2.82-5.03 (m, 7H), 1.83-1.95 (m, 1H), 1.69-1.80 (m, 2H),1.60 (s, 3H), 1.23-1.48 (m, 3H), 0.73-0.81 (m, 6H) 3-1-2 497.3 ¹H NMR(400 MHz, CHLOROFORM-d) δ 7.75 (br s, 1H), 7.43-7.57 (m, 2H), 7.26- 7.34(m, 1H), 7.18 (t, J = 9.12 Hz, 1H), 6.49-6.69 (m, 1H), 6.32-6.44 (m,1H), 5.78 (dd, J = 1.66, 10.57 Hz, 1H), 2.61-5.44 (m, 7H), 1.86-1.98 (m,1H), 1.62-1.85 (m, 2H), 1.59 (s, 3H), 1.36 (d, J = 6.84 Hz, 3H), 0.77(br d, J = 6.43 Hz, 6H)

For compounds in Table 5, in which a mixture of atropisomers andphosphorus isomers are listed, the following assay conditions wereemployed:

Coupled Nucleotide Exchange Assay:

Purified GDP-bound KRAS protein (aa 1-169), containing both G12C andC118A amino acid substitutions and an N-terminal His-tag, waspre-incubated in assay buffer (25 mM HEPES pH 7.4, 10 mM MgCl₂, and0.01% Triton X-100) with a compound dose-response titration for either 5min or 2 hours (see Table 15). Following compound pre-incubation,purified SOS protein (aa 564-1049) and GTP (Roche 10106399001) wereadded to the assay wells and incubated for an additional 30 min (for 5min compound pre-incubation) or 1 hour (for 2 hour compoundpre-incubation). To determine the extent of inhibition of SOS-mediatednucleotide exchange, purified GST-tagged cRAF (aa 1-149), nickel chelateAlphaLISA acceptor beads (PerkinElmer AL108R), and AlphaScreenglutathione donor beads (PerkinElmer 6765302) were added to the assaywells and incubated for 10 minutes. The assay plates were then read on aPerkinElmer EnVision Multilabel Reader, using AlphaScreen® technology,and data were analyzed using a 4-parameter logistic model to calculateIC₅₀ values.

Phospho-ERK1/2 MSD Assay:

MIA PaCa-2 (ATCC® CRL-1420™)-cells were cultured in RPMI 1640 Medium(ThermoFisher Scientific 11875093) containing 10% fetal bovine serum(ThermoFisher Scientific 16000044) and 1×penicillin-streptomycin-glutamine (ThermoFisher Scientific 10378016).Sixteen hours prior to compound treatment, MIA PaCa-2 cells were seededin 96-well cell culture plates at a density of 25,000 cells/well andincubated at 37° C., 5% CO₂. A compound dose-response titration wasdiluted in growth media, added to appropriate wells of a cell cultureplate, and then incubated at 37° C., 5% CO₂ for 2 or 4 hours. Followingcompound treatment, cells were stimulated with 10 ng/mL EGF (Roche11376454001) for 10 min, washed with ice-cold Dulbecco'sphosphate-buffered saline, no Ca²⁺ or Mg²⁺ (ThermoFisher Scientific14190144), and then lysed in RIPA buffer (50 mM Tris-HC pH 7.5, 1%Igepal, 0.5% sodium deoxycholate, 150 mM NaCl, and 0.5% sodium dodecylsulfate) containing protease inhibitors (Roche 4693132001) andphosphatase inhibitors (Roche 4906837001). Phosphorylation of ERK1/2 incompound-treated lysates was assayed using Phospho-ERK1/2 Whole CellLysate kits (Meso Scale Discovery KI51DWD) according to themanufacturer's protocol. Assay plates were read on a Meso ScaleDiscovery Sector Imager 6000, and data were analyzed using a 4-parameterlogistic model to calculate IC₅₀ values.

TABLE 5 Biochemical and Cellular Activity of Compounds (Mixture ofatropisomers and single isomers) Coupled Exchange p-ERK IC₅₀ (MIA- Ex. #IC₅₀ (μM) PaCa-2, μM) 1-1 2.68 1.23 1-1-1 250 10 1-1-2 1.43 0.59 1-220.1 1.63 1-2-1 5.15 1.45 1-2-2 >250 — 1-3 1.69 0.95 1-3-1 201 — 1-3-20.55 0.35 1-4 19.3 — 1-4-1 11.2 2.74 1-4-2 139 — 1-5 25.9 — 1-5-1 186 —1-5-2 14.9 — 1-6 7.28 3.31 1-7 8.04 1.76 1-7-1 5.28 1.72 1-7-2 7.74 3.261-8 5.78 3.69 1-8-1 53.8 — 1-8-2 4.95 1.62 2-1 18.6 — 2-2 3.37 1.77 3-114.3 — 3-1-1 94.7 — 3-1-2 13.3 1.77 (−) denotes not tested

The present invention is described in connection with preferredembodiments. However, it should be appreciated that the invention is notlimited to the disclosed embodiments. It is understood that, given thedescription of the embodiments of the invention herein, variousmodifications can be made by a person skilled in the art. Suchmodifications are encompassed by the claims below.

What is claimed is:
 1. A compound of having a structure of Formula I

wherein R¹ is a -C₁-C₆ alkyl or -C₃-C₆ cycloalkyl group; R^(1a) is a-C₁-C₆ heteroalkyl, aryl, heteroaryl, -C₃-C₆ cycloalkyl, or -C₃-C₆heterocycloalkyl group; or R¹ and R^(1a) together with the carbon atomto which they are attached, form a carbocyclic or heterocycloalkyl ring,wherein the carbocyclic or heterocycloalkyl ring can be unsubstituted orfused to an aromatic ring; R² is an aryl substituted with a halo, -OH,or NH_(2;) R³ is halo; R⁴ is H or methyl; R⁵ is H or methyl; or astereoisomer thereof, an atropisomer thereof, a pharmaceuticallyacceptable salt thereof, a pharmaceutically acceptable salt of thestereoisomer thereof, or a pharmaceutically acceptable salt of theatropisomer thereof.
 2. The compound of claim 1, wherein R¹ is an -C₁-C₆alkyl or -C₃-C₆ cycloalkyl group.
 3. The compound of claim 2, wherein R¹is methyl.
 4. The compound of claim 2, wherein R¹ is a cyclopropylgroup.
 5. The compound of claim 1, wherein R^(1a) is an -C₁-C₆ alkyl,aryl, or -C₃-C₆ cycloalkyl group.
 6. The compound of claim 5, whereinR^(1a) is an ethyl group.
 7. The compound of claim 5, wherein R^(1a)branched C₄ alkyl group.
 8. The compound of claim 5, wherein R^(1a) is acyclopropyl group.
 9. The compound of claim 5, wherein R_(1a) cyclobutylgroup.
 10. The compound of claim 5, wherein R^(1a) is a cyclopentylgroup.
 11. The compound of claim 5, wherein R^(1a) is a phenyl group.12. The compound of claim 1, wherein R¹ and R^(1a), together with thecarbon atom to which they are attached, form a 4-10 membered ring. 13.The compound of claim 12, wherein R¹ and R^(1a), together with thecarbon atom to which they are attached, form a cyclopentane.
 14. Thecompound of claim 12, wherein R¹ and R^(1a), together with the carbonatom to which they are attached, form a cyclohexane.
 15. The compound ofclaim 12, wherein R¹ and R^(1a) together with the carbon atom to whichthey are attached, form a 5-membered carbocyclic ring, wherein thecarbocyclic ring can be unsubstituted or fused to an aromatic ring. 16.The compound of claim 1, wherein R² is a fluorinated phenyl.
 17. Thecompound of claim 1, wherein R³ is Cl.
 18. The compound of claim 1,wherein R⁴ is H.
 19. The compound of claim 1, wherein R⁴ is methyl. 20.A compound, wherein the compound is selected from the group consistingof

or a stereoisomer thereof, an atropisomer thereof, a pharmaceuticallyacceptable salt thereof, a pharmaceutically acceptable salt of thestereoisomer thereof, or a pharmaceutically acceptable salt of theatropisomer thereof.
 21. A pharmaceutical composition comprising thecompound of claim 1 and a pharmaceutically acceptable excipient.
 22. Amethod of treating cancer in a subject in need thereof, the methodcomprising administering to the subject a therapeutically effectiveamount of the compound of claim 1, wherein the cancer is selected fromlung cancer and colorectal cancer.
 23. The method of claim 22, whereinthe cancer is lung cancer.
 24. The method of claim 22, wherein thecancer is colorectal cancer.
 25. The method of claim 23, wherein thelung cancer is non-small cell lung cancer.
 26. A pharmaceuticalcomposition comprising a compound of claim 20 and a pharmaceuticallyacceptable excipient.
 27. A method of treating cancer in a subject inneed thereof, the method comprising administering to the subject atherapeutically effective amount of the compound of claim 20, whereinthe cancer is selected from lung cancer and colorectal cancer.
 28. Themethod of claim 27, wherein the cancer is lung cancer.
 29. The method ofclaim 27, wherein the cancer is colorectal cancer.
 30. The method ofclaim 27, wherein the lung cancer is non-small cell lung cancer.